[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"cgt-wiki-knowledge-base":3,"header-xize-product-categories":364,"header-design-service-categories":534},{"entries":4,"categories":352},[5,14,20,26,32,38,47,53,59,65,71,77,83,89,98,107,113,119,125,131,136,142,148,154,160,166,172,178,184,190,196,202,208,214,220,226,232,237,243,249,255,259,265,274,280,286,292,298,304,310,316,322,328,334,340,346],{"id":6,"service_pic":7,"service_title":8,"service_class":6,"class_name":9,"service_content":10,"service_tag":11,"created_at":12,"updated_at":13},1,"","AAV structure and recombinant vector","AAV Knowledge Base","\u003Cp>AAV is a non-enveloped icosahedral parvovirus, about 25 nm. The capsid is composed of 60 VP subunits; the unique region of VP1 is involved in endosomal escape, and VP3 constitutes the main structural skeleton. The wild-type genome contains rep, cap and ITR at both ends; recombinant AAV usually retains only the ITR, and provides rep\u002Fcap and auxiliary functions in the production system, thereby separating the replication ability from the transgene expression cassette. [4-1 to 4-4]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028010\">\u003C\u002Fa>Function chain determined by structure\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"77\">\n\u003Cp>\u003Cstrong>Level\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"182\">\n\u003Cp>\u003Cstrong>Key components\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"336\">\n\u003Cp>\u003Cstrong>Direct impact on the experiment\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"77\">\n\u003Cp>Capsid\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"182\">\n\u003Cp>VP1\u002FVP2\u002FVP3 and surface variable regions\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"336\">\n\u003Cp>Receptor binding, cell entry, antibody recognition, tissue tropism and production of aptamers\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"77\">\n\u003Cp>Vector genome\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"182\">\n\u003Cp>ITR-promoter-transgene-regulatory element-polyA-ITR\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"336\">\n\u003Cp>Capacity, expression intensity, specificity, genome integrity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"77\">\n\u003Cp>Cell entrance\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"182\">\n\u003Cp>Glycans, AAVR and other dependent factors\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"336\">\n\u003Cp>The same capsid may produce different results in different cells and species\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"77\">\n\u003Cp>Production components\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"182\">\n\u003Cp>rep\u002Fcap, auxiliary genes, production cells\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"336\">\n\u003Cp>Yield, empty\u002Fsolid shell ratio, residue and batch-to-batch consistency\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>The 2023 biophysical review emphasized that transduction is a continuous process of receptor binding, endocytosis, endosomal escape, nuclear transport, uncoating, and second-strand formation; the structure of the Rep packaging complex in 2026 further showed that packaging is not a simple \"DNA loading into an empty shell\", but a dynamic process of collaboration between Rep, ITR and capsid interface. [4-3,4-4]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028011\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[4-1] Pillay S, et al. An essential receptor for adeno-associated virus infection. Nature. 2016;530:108-112. DOI:10.1038\u002Fnature16465. PMID:26814968. [H]\u003C\u002Fp>\n\u003Cp>[4-2] Bennett A, Mietzsch M, Agbandje-McKenna M. Understanding capsid assembly and genome packaging for adeno-associated viruses. Future Virol. 2017;12:283-297. DOI:10.2217\u002Ffvl-2017-0011. PMID:36776482. [H]\u003C\u002Fp>\n\u003Cp>[4-3] Xiang YS, Hao GG. Biophysical characterization of adeno-associated virus capsid through the viral transduction life cycle. Biophys Rev. 2023;21:62. DOI:10.1186\u002Fs43141-023-00518-5. PMID:37195476. [N]\u003C\u002Fp>\n\u003Cp>[4-4] Xu V, et al. Structural basis for Rep-mediated adeno-associated virus packaging. Cell Rep. 2026;45:117044. DOI:10.1016\u002Fj.celrep.2026.117044. PMID:41790558. [N]\u003C\u002Fp>","AAV, AAV Knowledge Base","2026-09-18 10:53:25","2026-10-28 22:43:53",{"id":15,"service_pic":7,"service_title":16,"service_class":6,"class_name":9,"service_content":17,"service_tag":11,"created_at":18,"updated_at":19},2,"ssAAV, scAAV and packaging capacity","\u003Cp>ssAAV needs to complete complementary strand synthesis in the cell before forming a transcription template; scAAV folds to form a double-stranded structure, which usually works faster and is more stable, but the available expression cassette capacity is approximately halved. Conventional ssAAV design should calculate the ITR at both ends within a total load of about 4.7 kb; the non-ITR expression cassette of scAAV is usually controlled at about 2.2-2.4 kb. As the upper limit is approached, not only may yields decrease, but the proportion of truncated or heterogeneous genomes may increase. [5-1 to 5-4]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028013\">\u003C\u002Fa> Processing sequence of ultra-long expression cassettes\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>\u003Cstrong>Sequence\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>\u003Cstrong>Processing actions\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>\u003Cstrong>Judgment Criteria\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>Recalculate the complete load\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>All sequences between ITRs are counted; confirm promoter, intron, WPRE, tag and polyA\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>2\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>Compression control element\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>Prioritize to use a validated short promoter, short polyA or delete unnecessary tags\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>3\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>Replacement of compact functional components\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>Use shorter coding sequences, split editors, or more compact control systems\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>4\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>Adopt dual AAV\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>Clear the recombination mechanism, linker design, double-vector ratio and co-transduction bottleneck\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"53\">\n\u003Cp>5\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>Change delivery platform\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"350\">\n\u003Cp>When dual carriers cannot meet efficiency or consistency, overload packaging is no longer the default solution\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>\"Ability to detect packaging\" does not equate to \"resulting in a product that is uniform, complete and has the intended efficacy.\" Overloading vectors should be assessed for genomic integrity using long-read sequencing or orthogonal methods covering the entire expression cassette. [5-3]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028014\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[5-1] McCarty DM. Self-complementary AAV vectors: advances and applications. Mol Ther. 2008;16:1648-1656. DOI:10.1038\u002Fmt.2008.171. [F]\u003C\u002Fp>\n\u003Cp>[5-2] Bennett A, et al. Understanding capsid assembly and genome packaging for adeno-associated viruses. Future Virol. 2017;12:283-297. DOI:10.2217\u002Ffvl-2017-0011. [H]\u003C\u002Fp>\n\u003Cp>[5-3] Tran NT, et al. AAV-genome population sequencing of vectors packaging CRISPR components reveals design-influenced heterogeneity. Mol Ther Methods Clin Dev. 2020;18:639-651. DOI:10.1016\u002Fj.omtm.2020.07.007. PMID:32775498. [H]\u003C\u002Fp>\n\u003Cp>[5-4] Davis JR, et al. Efficient prime editing in mouse brain, liver and heart with dual AAVs. Nat Biotechnol. 2024;42:253-264. DOI:10.1038\u002Fs41587-023-01758-z. PMID:37142705. [N]\u003C\u002Fp>","2026-09-18 10:53:50","2026-10-16 21:53:42",{"id":21,"service_pic":7,"service_title":22,"service_class":6,"class_name":9,"service_content":23,"service_tag":11,"created_at":24,"updated_at":25},3,"Titer, dose and particle quality","\u003Cp>Titer is not a single indicator. vg\u002FmL describes the amplifiable vector genomic signal, capsid particles\u002FmL describes total capsids, and infectivity or transduction units reflect the functional output in a specific system. Different primers, standards, lysis and cell-free DNA removal conditions will change the qPCR\u002FddPCR results, so cross-laboratory comparisons should first check methods instead of just comparing numbers. [6-1 to 6-5]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028016\">\u003C\u002Fa>Explanation of three levels of quality\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"96\">\n\u003Cp>\u003Cstrong>Level\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>\u003Cstrong>Representative indicators\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"288\">\n\u003Cp>\u003Cstrong>Questions that cannot be answered individually\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"96\">\n\u003Cp>Quantity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>vg\u002FmL, total vg, total capsids\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"288\">\n\u003Cp>Whether the genome is complete and whether the particles can be transduced\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"96\">\n\u003Cp>Composition\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Empty\u002Ffull shell, partially loaded, aggregate, residual DNA\u002Fprotein\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"288\">\n\u003Cp>Expression efficacy in target cells\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"96\">\n\u003Cp>Function\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Infectivity, transduction efficiency, transgene expression or biological activity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"288\">\n\u003Cp>Direct comparability across cell lines and species\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>The empty\u002Freal shell ratio should be interpreted by AUC, AEX, mass spectrometry, or other orthogonal means; \"real shells\" may still contain partial genomes or anomalous fragments. Research in 2023-2024 will continue to promote multi-attribute detection of genome integrity and capsid identity. Therefore, QC configuration should be based on uses and risks rather than pursuing the optimization of a single indicator. [6-3 to 6-5]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028017\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[6-1] Gimpel AL, et al. Analytical methods for process and product characterization of recombinant AAV-based gene therapies. Mol Ther Methods Clin Dev. 2021;20:740-754. DOI:10.1016\u002Fj.omtm.2021.02.010. PMID:33738328. [H]\u003C\u002Fp>\n\u003Cp>[6-2] Werle AK, et al. Comparison of analytical techniques to quantitate the capsid content of AAV vectors. Mol Ther Methods Clin Dev. 2021;23:254-262. DOI:10.1016\u002Fj.omtm.2021.08.009. PMID:34703846. [H]\u003C\u002Fp>\n\u003Cp>[6-3] Richter K, et al. Purity and DNA content of AAV capsids assessed by analytical ultracentrifugation and orthogonal biophysical techniques. Eur J Pharm Biopharm. 2023. PMID:37196871. [N]\u003C\u002Fp>\n\u003Cp>[6-4] Smith J, et al. Rapid AAV identity testing through intact mass analysis of full-length capsid proteins. J Proteome Res. 2023. DOI:10.1021\u002Facs.jproteome.3c00513. PMID:38123456. [N]\u003C\u002Fp>\n\u003Cp>[6-5] Park S, et al. Enhancing AAV2 and AAV9 production with high full-capsid ratio by design-of-experiment optimization. Biotechnol J. 2024;19:e2300667. DOI:10.1002\u002Fbiot.202300667. PMID:38479987. [N]\u003C\u002Fp>","2026-09-18 10:54:16","2026-10-16 21:26:42",{"id":27,"service_pic":7,"service_title":28,"service_class":6,"class_name":9,"service_content":29,"service_tag":11,"created_at":30,"updated_at":31},4,"AAV advantages, limitations and immune factors","\u003Ch2>\u003Ca name=\"_Toc240028019\">\u003C\u002Fa>Common advantages\u003C\u002Fh2>\n\u003Cul>\n\u003Cli>Multiple levels of targeting can be achieved through natural or engineered capsids and transcriptional regulation; longer-term expression can be achieved in many terminally differentiated cells. \u003C\u002Fli>\n\u003Cli>Recombinant vectors usually exist in extrachromosomal form and are suitable for in vivo delivery of secreted proteins, enzymes, reporter genes, regulatory elements and partial editing systems. \u003C\u002Fli>\n\u003Cli>Existing clinical experience in the eye, liver, muscle and nervous system provides a mature analysis and risk framework for study design. \u003C\u002Fli>\n\u003C\u002Ful>\n\u003Ch2>\u003Ca name=\"_Toc240028020\">\u003C\u002Fa>Limitations that must be proactively prompted\u003C\u002Fh2>\n\u003Cul>\n\u003Cli>Packaging capacity is limited; scAAV capacity is smaller, and dual AAV will introduce co-transduction and recombination efficiency issues. \u003C\u002Fli>\n\u003Cli>Neutralizing antibodies formed from previous exposure may reduce transduction and limit repeated dosing; both capsids and transgenes can trigger innate or adaptive immunity. \u003C\u002Fli>\n\u003Cli>High systemic doses are associated with hepatotoxicity, complement-related events, and thrombotic microangiopathy; DRG pathology is seen in NHP with CSF or systemic administration. \u003C\u002Fli>\n\u003Cli>Duration of expression depends on cell turnover, immune response, promoter silencing and disease context and should not be uniformly committed. \u003C\u002Fli>\n\u003Cli>Mouse results cannot be directly extrapolated to NHPs or humans; receptor differences can create significant species barriers for engineered capsids. \u003C\u002Fli>\n\u003C\u002Ful>\n\u003Ch2>\u003Ca name=\"_Toc240028021\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[7-1] Li C, Samulski RJ. Engineering adeno-associated virus vectors for gene therapy. Nat Rev Genet. 2020;21:255-272. DOI:10.1038\u002Fs41576-019-0205-4. PMID:32042148. [H]\u003C\u002Fp>\n\u003Cp>[7-2] Costa Verdera H, Kuranda K, Mingozzi F. AAV vector immunogenicity in humans. Mol Ther. 2020;28:723-746. DOI:10.1016\u002Fj.ymthe.2019.12.010. PMID:31972133. [H]\u003C\u002Fp>\n\u003Cp>[7-3] Muhuri M, et al. Overcoming innate immune barriers that impede AAV gene therapy vectors. J Clin Invest. 2021;131:143780. DOI:10.1172\u002FJCI143780. PMID:33393506. [H]\u003C\u002Fp>\n\u003Cp>[7-4] Hordeaux J, et al. AAV-induced dorsal root ganglion pathology. Hum Gene Ther. 2020;31:808-818. DOI:10.1089\u002Fhum.2020.167. PMID:32845779. [H]\u003C\u002Fp>\n\u003Cp>[7-5] Lek A, et al. Death after high-dose rAAV9 gene therapy in Duchenne muscular dystrophy. N Engl J Med. 2023;389:1203-1210. DOI:10.1056\u002FNEJMoa2307798. PMID:37754285. [N]\u003C\u002Fp>","2026-09-18 10:54:51","2026-10-19 13:54:52",{"id":33,"service_pic":7,"service_title":34,"service_class":6,"class_name":9,"service_content":35,"service_tag":11,"created_at":36,"updated_at":37},5,"Selecting Vector Functions by Research Objective","\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>\u003Cstrong>Research Purpose\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>\u003Cstrong>Expression box core\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>\u003Cstrong>Primary Risks\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Overexpression\u002FFunctional Rescue\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>Coding sequence + adapter promoter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>Overexpression toxicity, non-target tissue expression\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>RNA intervention\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>shRNA\u002FmiRNA or regulatory RNA\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>Dose-dependent toxicity, off-target and processing saturation\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Gene editing\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>Nuclease\u002FEditor + gRNA; double AAV if necessary\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>Capacity, immunity, off-target, long-term nuclease expression\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Neural circuit tracing\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>fluorescence\u002Ftranssynaptic or retrograde tools\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>Directivity, leakage, toxicity and time windows\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Sensing and Control\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"230\">\n\u003Cp>Sensors, DREADD\u002Foptogenetic components\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"250\">\n\u003Cp>Expression level affects signal-to-noise ratio and physiological status\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>The selection sequence should first determine \"what the cell needs to do\" and then decide on the capsid. The capsid mainly affects arrival and entry, and the promoter\u002Fenhancer determines the expression profile after entry, and the two cannot replace each other. Editing and tracing projects should also set up independent controls for the vector itself, effectors, and readout systems. [9-1 to 9-3]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028025\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[9-1] Wang D, Zhang F, Gao G. CRISPR-based therapeutic genome editing: strategies and in vivo delivery by AAV vectors. Cell. 2020;181:136-150. DOI:10.1016\u002Fj.cell.2020.03.023. PMID:32243786. [H]\u003C\u002Fp>\n\u003Cp>[9-2] Tervo DGR, et al. A designer AAV variant permits efficient retrograde access to projection neurons. Neuron. 2016;92:372-382. DOI:10.1016\u002Fj.neuron.2016.09.021. PMID:27720486. [H]\u003C\u002Fp>\n\u003Cp>[9-3] Davis JR, et al. Efficient prime editing in mouse brain, liver and heart with dual AAVs. Nat Biotechnol. 2024;42:253-264. DOI:10.1038\u002Fs41587-023-01758-z. [N]\u003C\u002Fp>","2026-09-18 10:56:00","2026-10-16 21:21:28",{"id":39,"service_pic":7,"service_title":40,"service_class":41,"class_name":42,"service_content":43,"service_tag":44,"created_at":45,"updated_at":46},7,"Molecular basis of tropism",6,"Six Variables for Serotype Selection","\u003Cp>\u003Cstrong>The concept of two-tiered events. \u003C\u002Fstrong>AAV's preference (tropism) for a certain tissue\u002Fcell is not a single event, but is determined by two layers of molecular events: \"attachment receptor\" and \"required entry receptor\". Confusing these two layers is the source of misjudgment of tissue vg as target cell expression. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Attachment factor:\u003C\u002Fstrong> Glycosaminoglycans\u002Fglycoproteins determine the initial binding affinity and tissue distribution tendency of the capsid and the cell surface. Most of them are glycans (HSPG, sialic acid, galactose). \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Essential entry receptor:\u003C\u002Fstrong> Protein factor that determines endocytosis, reverse vesicle transport (TGN orientation), and infection success or failure. AAVR (KIAA0319L) is the broadest spectrum essential entry receptor discovered so far [10-5]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Key corollary. \u003C\u002Fstrong>Just because a cell has a certain serotype of attached glycan on its surface does not mean that the cell can be effectively transduced; it must also express AAVR (or its alternative receptor such as CPD) and subsequent trafficking factors. This explains why \"high tissue vg\" does not equal \"high target cell expression\". \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Receptor\u002Fco-receptor × serotype correspondence summary table. \u003C\u002Fstrong>The corresponding relationships between receptors\u002Fco-receptors for commonly used capsids are as follows. \u003C\u002Fp>\n\u003Ctable>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>Serotype\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>Main attachment receptor\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>Required to enter the receptor\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>Reported co-receptors (mostly not verified by genetic screening)\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV1\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>α2-3 N-linked sialic acid[10-6][10-7]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Unknown\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV2\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>HSPG (basic residues such as R585\u002FR588\u002FK532) [10-8]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>FGFR1, αVβ5\u002Fα5β1 integrin, HGFR, LamR, CD9 (knockout screening did not confirm essential) [10-9]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV3B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>HSPG (weaker, R594 mediated)[10-10]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>FGFR1, HGFR, LamR[10-9]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV4\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>α2-3 O-linked sialic acid (mucin-type O-glycan)[10-6][10-11]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR optional (Clade G, can bypass AAVR) [10-12]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Unknown\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV5\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>α2-3 N-linked sialic acid[10-6][10-13]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>PDGFRα\u002FPDGFRβ[10-14]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV6\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>HSPG + α2-3 and α2-6 N-linked sialic acid (double linkage)[10-7][10-15]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>EGFR[10-16]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV7\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Not confirmed (glycan phenotype is close to the AAV8\u002F9 branch)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Unknown\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV8\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>LamR (reported); there is also CPD\u002FAAVR2 as an alternative essential receptor [10-17]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR\u002FCPD[10-5][10-17]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Unknown\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV9\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Terminal N-linked galactose (asialo exposed)[10-18][10-19]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>LamR, putative integrin[10-9]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAVrh.10\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Sulfated N-acetyllactosamine (sulfated LacNAc, terminal galactose epitope) [10-9]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR[10-5]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Unknown\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAVrh74\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Highly homologous to AAV8 (Clade E); the glycan profile is similar to AAV8, and has not been accurately mapped separately (to be verified)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Inferred AAVR (to be verified)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV-DJ\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>HSPG (chimera, derived from AAV2\u002F8\u002F9)[10-20]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>See chimeric sources\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"165\">\n\u003Cp>\u003Cstrong>AAV-PHP.B\u002FPHP.eB\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>Retains AAV9 galactose binding; inserted peptide binds Ly6A\u002FSca-1 (GPI-anchored protein) as BBB crossing receptor[10-21][10-22]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>AAVR + Ly6A\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"165\">\n\u003Cp>—\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp>The classic hemagglutination experiment of Kaludov et al. 2001 proved that AAV4 uses α2-3 O-linked sialic acid (not α2-6), and AAV5 uses α2-3 N-linked sialic acid (not α2-6); it is AAV6 that truly recognizes both α2-3 and α2-6 N-links, and AAV1 only recognizes α2-3 N-links [10-6][10-7]. This difference determines that AAV4\u002F5 prefers epithelia with high O-glucose\u002FN-glucose abundance, such as the airway\u002Fependymum, while AAV6 has a wider double junction spectrum and stronger airway transduction. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAVR\u003C\u002Fstrong>\u003Cstrong>(KIAA0319L):\u003C\u002Fstrong>Type I transmembrane protein, extracellular contains MANSC domain and multiple Ig-like PKD domains. It binds to the capsid through the PKD2 domain, is rapidly endocytosed, and transported to the trans-Golgi network (TGN) [10-5]. Most natural and engineered capsids, such as AAV1\u002F2\u002F3B\u002F5\u002F6\u002F8\u002F9\u002Frh.10, rely on AAVR; exceptions are AAV4 and the closely related Clade G that can bypass AAVR [10-12]. In 2025, CD164 was also identified as an essential receptor for divergent AAVs [10-23]. AAVR is widely expressed in most nucleated cells, so it does not constitute a major determinant of tissue tropism. The difference in tropism is mainly determined by the upper attachment glycans\u002Fco-receptors; forced expression of AAVR in a certain tissue can make originally low-transducing tissues transduced by broad-spectrum AAV [10-24]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>HSPG (heparan sulfate proteoglycan):\u003C\u002Fstrong>Summerford and Samulski established it as the main receptor for AAV2 in 1998, with key residues R585\u002FR588 (and R484\u002FR487\u002FK532) [10-8]. Those with HSPG as the main attachment receptor are AAV2, AAV3B, AAV6, and AAV13 [10-9]. Recent evidence suggests that HSPG binding of native AAV2 clinical strains is not strong, and that HSPG binding may be a culture-adaptive trait acquired by in vitro passage. HSPG is widely present in vascular endothelium, liver sinusoids, and renal tubular basement membranes, making AAV2 enriched in the liver after systemic administration, but it is also easily intercepted by liver non-parenchymal cells [10-9]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Sialic acid (the connection method determines the subdivision):\u003C\u002Fstrong>α2-3 N-linked is AAV1, AAV5[10-6][10-13]; α2-3 O-linked is AAV4 (mucin type) [10-6]; α2-3+α2-6 N-linked is AAV6[10-7]. The high abundance of sialic acid on the airway\u002Falveolar epithelial surface makes AAV4\u002F5\u002F6 bias toward the lung; the difference in glycoforms between the cerebral ependymium and RPE determines that AAV4 is biased towards the membrane and AAV5 is biased towards the glial. Deletion of the CMAH enzyme brings similar sialic acid profiles between humans and mice, which is one of the reasons why SIA-dependent capsids are relatively conserved across species [10-9]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Galactose (the organizational key of AAV9\u002Frh.10):\u003C\u002Fstrong>AAV9 recognizes the terminal N-linked galactose exposed after desialylation, and the key residues N470\u002FD271\u002FN272\u002FY446\u002FW503 constitute the base pocket of the tripodal protrusion [10-18][10-19]; AAVrh.10 recognizes sulfation LacNAc (also contains terminal galactose) [10-9]. Galactose exposure on the surface of neonates and muscle\u002Fmyocardial\u002Fpulmonary epithelial cells is high, which is the glycosyl base for AAV9 to cross muscle fibers and the BBB (neonatal); adult mice have low BBB endothelial galactose exposure, so AAV9 has weak adult BBB crossing [10-25]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Other receptors\u002Fco-receptors:\u003C\u002Fstrong>PDGFRα\u002Fβ are AAV5 co-receptors and are expressed in oligodendrocyte precursors, vascular pericytes, and some neurons [10-14]; EGFR is an AAV6 co-receptor [10-16]. Although there were early reports of HGFR(c-Met)\u002FFGFR1\u002FLamR and integrin αVβ5\u002Fα5β1, multiple genome-wide CRISPR screens did not confirm that they are essential and should be regarded as “reported but not confirmed” [10-9]. CPD (carboxypeptidase D, also known as AAVR2) is an alternative essential receptor for AAV discovered in 2025 and is important for AAV8 and AAV11 [10-17]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Mechanism summary. \u003C\u002Fstrong>Receptor expression differences determine affinity at four levels: ① Cell type level - hepatocytes vs Kupffer cells vs liver sinusoidal endothelium in the same organ have different AAVR, HSPG, and galactose exposures, resulting in \"organ enrichment ≠ target cell enrichment\"; ② Developmental level - newborn mice have higher exposure to galactose on the BBB and muscle cell surfaces than adults, and AAV9 neonatal administration CNS\u002Fmuscle transduction is much stronger than in adults [10-25]; ③ Species\u002Fstrain level - glycosyl modifying enzyme (CMAH), GPI-anchored protein (Ly6A), TfR1 sequence differences lead to cross-species tropism drift (see 6.3); ④ Epithelial polarity level - airway luminal surface vs basolateral surface, subretinal space vs vitreous cavity, receptor accessibility is completely different, and the route of administration therefore reshapes tropism (see 6.3) 6.4). \u003C\u002Fp>","AAV, Six Variables for Serotype Selection","2026-09-18 10:58:47","2026-10-16 22:23:16",{"id":48,"service_pic":7,"service_title":49,"service_class":41,"class_name":42,"service_content":50,"service_tag":44,"created_at":51,"updated_at":52},8,"Native Serotype × Tissue Tropism Matrix","\u003Cp>The table below summarizes the relative consensus transduction potency (high\u002Fmoderate\u002Flow\u002Fvery low\u002Fnone) of 14 commonly used capsids upon IV systemic administration in adult C57BL\u002F6 mice, along with representative quantitative data and notes on route of administration. Because there are too many tissue columns in 14 capsids × 17, it is split into three sub-tables based on system periphery, CNS and retina, and lung\u002Fskin\u002Fbone. Key sources: Zincarelli 2008 (IV mouse types 1-9 head-to-head) [10-26]; Yang et al. 2024 (21 capsid barcode library, C57BL\u002F6\u002FBALB\u002Fc\u002Fcynomolgus monkey IV, 5×10¹² vg\u002Fkg) [10-27]; Bish 2008 (heart) [10-28]; Halbert 2001 (Lung) [10-29]; Chan 2017 (PHP.eB) [10-21]. Intensity is a relative qualitative judgment, with quantitative\u002Fpathway remarks in parentheses, which cannot be directly extrapolated across studies; \u003Cbr \u002F>Sub-table A | System and peripheral tissue (IV mainly throughout the body)\u003C\u002Fp>\n\u003Ctable>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Capsid\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Liver (liver cells)\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Skeletal Muscle\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Myocardium\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Kidney\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Pancreas\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Fat\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>Tumor\u002FEndothelial\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV1\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Mid (IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV\u002FIM)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV)[10-28]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV2\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, but lower than 8\u002F9)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low-Medium (IM Medium)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low-Medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium (partial)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium (tumor model)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV3B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (hepatic hematopoietic progenitor cell preference)[10-9]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (HCC\u002Fhematopoietic)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV4\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low (off-target in liver, consistent across species)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (glomerulus)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (islet)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV5\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium(IM)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV6\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium-High (IV)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium-high (IM, may be accompanied by myositis)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (excellent system heart) [10-34]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium (acinus)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV7\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, high expression group)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV)[10-28]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV8\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>One of the highest (IV, conserved across species)[10-26][10-35]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV\u002FIM)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, slightly lower than 9)[10-28]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium-High[10-35]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium (acinus)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV9\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, liver is still the largest reservoir)[10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, whole body muscle fiber) [10-26]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Highest (IV, global myocardium better than 1\u002F6\u002F7\u002F8)[10-28]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (gastrointestinal\u002Fpancreatic)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (brown fat)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAVrh.10\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Highest (consistent across rats\u002Fmonkeys) [10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, slightly lower than 9) [10-36]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAVrh74\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (near AAV8, effective across rat\u002Fmonkey liver)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Highest (IV, Elevidys clinical use)[10-37][10-38]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (IV, slightly lower than 9) [10-36]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low-Medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV-DJ\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>High (mouse liver≈AAV8\u002F9)[10-20]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium-High\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV-PHP.B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium (CNS\u002Fhigh liver disease)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low-Medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"82\">\n\u003Cp>\u003Cstrong>AAV-PHP.eB\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Medium-high (IV, liver is still high)[10-21]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"82\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp>Sub-table B｜Central Nervous System and Retina\u003C\u002Fp>\n\u003Ctable>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>Capsid\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>CNS-Neuron\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>CNS-Astrocyte\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>Cerebrovascular\u002FEndothelial\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>Retina-RPE\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>Retina-Photoreceptor\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>Retina-Ganglia\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV1\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (ICV\u002Flocal; IV low)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium(ICV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)[10-30]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)[10-30]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV2\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (local injection; very low IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (IVT biased toward RGC)[10-31]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium(SR)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (IVT classic)[10-31]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV3B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV4\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low (IV); tubular\u002Fsubventricular zone high (ICV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium (ICV deflection membrane)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR, RPE specific)[10-30]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low-medium (SR)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV5\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium-High (partial, Purkinje, etc.)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (colloid preference)[10-32]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)[10-30]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)[10-30]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV6\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low(IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV7\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low(IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV8\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low (IV trace; locally high)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)[10-31]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR, stronger than AAV2)[10-31]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV9\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium (IV; high neuron in newborns, weak in adults)[10-25]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium (adult partial gelatin)[10-25]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low (IV is weak across BBB)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium-High (SR)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (SR)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Medium(IVT)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAVrh.10\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Mid (IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low-Medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAVrh74\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low-Medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV-DJ\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low(IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (RPE preference)[10-39]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV-PHP.B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>The highest (IV, only C57BL\u002F6 series, > dozens of times AAV9) [10-22]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (IV, gelatinous)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (brain endothelial crossing)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"94\">\n\u003Cp>\u003Cstrong>AAV-PHP.eB\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Highest (IV, 1×10¹¹vg transduces 69% cortical\u002F55% striatal neurons)[10-21]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (IV)[10-21]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>High (brain endothelium)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"94\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp>Sublist C｜Lung, skin, bone\u002Fcartilage\u003C\u002Fp>\n\u003Ctable>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>Capsid\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>Lung-airway epithelium\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>Lung-Alveoli\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>Skin\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>Bone\u002FCartilage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV1\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV2\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low (poor IV; partially weaker than 6)[10-29]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV3B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV4\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>High (local; alveolar 21.2% vs AAV9 1.7%)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>High (IV, lung tdTomato⁺~21%)[10-27]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV5\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>High (strong intracavitary administration)[10-29]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV6\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Highest (airway epithelium, about 3 times stronger than AAV2)[10-29]\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV7\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV8\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV9\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>medium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAVrh.10\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAVrh74\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV-DJ\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV-PHP.B\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"132\">\n\u003Cp>\u003Cstrong>AAV-PHP.eB\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"132\">\n\u003Cp>Low\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp>\u003Cstrong>Matrix notes. \u003C\u002Fstrong>The high CNS transduction of AAV-PHP.B\u002FPHP.eB is strictly limited to Ly6A-permissive strains such as C57BL\u002F6J (see 6.3), and this advantage disappears in BALB\u002Fc, CD-1, and NHP; the high CNS\u002Fretinal transduction of AAV2 is almost entirely from local injection (parenchyma, ICV, IVT), and its IV systemic administration has extremely low CNS transduction. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV1: \u003C\u002Fstrong>Muscle\u002Fnerve excellent. IM is highly efficient in skeletal muscle, ICV\u002Fbrain parenchymal neurons and ependyma are strong, SR administration is efficient in RPE and photosensitivity, IV is liver-enriched, etc.; it is often used for local administration of neuromuscular diseases and CNS [10-26][10-28][10-30]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV2\u003C\u002Fstrong>\u003Cstrong>: \u003C\u002Fstrong>The most well-studied, “moderately broad-spectrum but not extreme” affinity. Local brain injection and IVT retinal RGC are classic scenarios; IV liver enrichment but less than 8\u002F9, HSPG dependence makes it easy to be cleared by liver non-parenchymal cells combined with heparin [10-8][10-26]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV3B:\u003C\u002Fstrong> Liver and hematopoietic\u002Fhepatocellular carcinoma bias, recognizes HSPG+HGFR; used in hepatic progenitor cells and HCC models, systemic CNS\u002Fmuscle weakness [10-9]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV4:\u003C\u002Fstrong> Consistent liver off-target + lung\u002Falveolar\u002Fglomerular\u002Fislet preference across species; Yang 2024 showed AAV4 lung tdTomato⁺ 21.2% vs AAV9 1.7%, and this liver off-target is also true in cynomolgus monkeys [10-27]. ICV is in the ependyma, SR administration is specific to RPE (RPE65 has been used clinically); AAVR is not necessary, making it a framework for engineering low-hepatotoxicity vectors [10-12]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV5:\u003C\u002Fstrong>N-linked α2-3 sialic acid+PDGFR. ICV\u002Fbrain parenchyma is biased toward astrocytes, has high efficiency in airway luminal delivery, and strong photosensitivity\u002FRPE in SR delivery (AAV5 used in Spark\u002FRPGR clinical trials) [10-14][10-29][10-32]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV6\u003C\u002Fstrong>\u003Cstrong>:\u003C\u002Fstrong>The only one that simultaneously binds α2-3\u002Fα2-6 sialic acid + HSPG; optimal for airway epithelium (about 3 times stronger than AAV2) [10-29], excellent for IV systemic heart [10-34], and IM skeletal muscle can be accompanied by inflammatory response [10-33]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV7\u003C\u002Fstrong>\u003Cstrong>: \u003C\u002Fstrong>IV high expression group (same level as AAV9), strong in muscle\u002Fliver\u002Fheart, and low immunogenicity [10-26]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV8\u003C\u002Fstrong>\u003Cstrong>:\u003C\u002Fstrong> One of the gold standards for liver targeting, IV hepatocytes are highly efficient and relatively conserved across species [10-26][10-35]; SR administration is more photosensitive than AAV2 [10-31]; muscle, heart, kidney, and pancreas can all be transduced. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV9\u003C\u002Fstrong>\u003Cstrong>:\u003C\u002Fstrong> Broad spectrum + cross-BBB (strong in newborns, weak in adults) + strongest in global myocardium[10-25][10-28]. IV is high in skeletal muscle, cardiac muscle, and liver; IV in adults is weak across the BBB; SR is acceptable in all layers of the retina. Zolgensma (SMA) uses AAV9. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAVrh.10:\u003C\u002Fstrong>Isolated from rhesus monkeys, it recognizes sulfated LacNAc; the liver is consistently efficient across mice\u002Fmonkeys[10-27], and the muscle\u002Fheart is high, but the measured myocardial transduction of NHP is slightly lower than that of AAV9[10-36]; MPS IIIA and other clinical uses. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAVrh74:\u003C\u002Fstrong> Closely related to AAV8 (Clade E), muscle-biased; Elevidys is used for DMD, the MHCK7 promoter drives microdystrophin [10-37][10-38]; NHP is effective in both liver and muscle [10-27]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV-DJ\u003C\u002Fstrong>\u003Cstrong>:\u003C\u002Fstrong>AAV2\u002F8\u002F9 in vitro reassortment chimera, combined with HSPG; mouse liver ≈AAV8\u002F9 and transduces a variety of human cells better than the parent, with RPE preference [10-39]. Note: Liver dominance is only in mice, NHP is not conserved [10-27]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV-PHP.B\u003C\u002Fstrong>\u003Cstrong>:\u003C\u002Fstrong>AAV9 VR-IV inserts TLAVPFK peptide and binds Ly6A; IV extensively transduces C57BL\u002F6J brain parenchymal neurons + glia, still high in liver, only effective in C57BL\u002F6 line [10-22]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>AAV-PHP.eB:\u003C\u002Fstrong> Optimized version of PHP.B, 1×10¹¹ vg can transduce 69% cortex\u002F55% striatal neurons, the dose can be reduced to 1\u002F20 of PHP.B, partial to inhibitory neurons; still relies on Ly6A, strain restrictions are the same as PHP.B[10-21]. \u003C\u002Fp>","2026-09-18 11:13:31","2026-10-16 21:45:53",{"id":54,"service_pic":7,"service_title":55,"service_class":41,"class_name":42,"service_content":56,"service_tag":44,"created_at":57,"updated_at":58},9,"Species and Strain Differences in Tropism","\u003Cp>\u003Cstrong>PHP.B\u002FLy6A: Why only C57BL\u002F6J works. \u003C\u002Fstrong>Deverman et al. 2016 used the CREATES method to screen out AAV-PHP.B, IV to achieve widespread transduction in the whole brain in C57BL\u002F6J [10-22]. Hordeaux et al. 2018 directly proved that its neurotropism is limited to C57BL\u002F6J, and the advantage in the brain disappears in BALB\u002Fc, CD-1 and NHP; Batista et al. subsequently proved that FVB and DBA\u002F2 are partially allowed, but BALB\u002Fc and CBA\u002FJ are not allowed [10-40][10-41]. Mechanistically, Ly6A\u002FSca-1 (lymphocyte activation antigen 6A, GPI-anchored protein) is a receptor for PHP.B that crosses the BBB. It is highly expressed on the luminal surface of brain microvascular endothelium in C57BL\u002F6J and is not expressed in BALB\u002FcJ due to promoter polymorphism [10-42]. Cynomolgus\u002Frhesus monkeys do not have an ortholog of Ly6A, so PHP.B\u002FPHP.eB is not superior to AAV9 in NHP brains [10-27]. Conclusions: The PHP.B family is a mouse strain-specific research tool and cannot be extrapolated to NHP\u002Fhumans. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Humanized receptor designed capsid BI-hTFR1. \u003C\u002Fstrong>Huang et al. 2024 screened BI-hTFR1 through directed evolution, which binds to the luminal epitope of human transferrin receptor 1 (TfR1\u002FTFRC) (does not compete with endogenous transferrin ligand) [10-43]. It provides a 40-50-fold CNS reporter gene enhancement over AAV9 in human TFRC knock-in (KI) mice and is CNS-specific; this advantage is completely lost in wild-type (murine TfR1) mice. BI-hTFR1 does not recognize mouse TfR1 and must be evaluated in human TfR1 KI mice, human BBB models or NHP, and ordinary C57BL\u002F6 is ineffective - this is a paradigm case of \"receptor species determines capsid effectiveness\" [10-43]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Conservative list. \u003C\u002Fstrong>Relatively conservative and can be extrapolated across species: liver targeting of AAV8\u002FAAVrh.10\u002FAAVrh74 (both mice and cynomolgus monkeys have the most efficient capsid in the liver) [10-27][10-35]; AAV9 has a broad spectrum of cardiac\u002Fskeletal muscle (effective in mice, rats, dogs, and NHPs) [10-28]; AAV4 has liver off-targeting + lung\u002Fglomerular\u002Fislet bias (cross-targeting) Consistent with C57BL\u002F6, BALB\u002Fc, and cynomolgus monkeys) [10-27]; MyoAAV muscle preference (both mice and NHP show 10-50 times the muscle mRNA of the native type) [10-44]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Not a conservative list. \u003C\u002Fstrong>The differences are huge and cannot be directly extrapolated: BBB crossing of AAV-PHP.B\u002FPHP.eB\u002FCAP-B10 (only C57BL\u002F6 line, NHP has no advantage) [10-27][10-40]; liver advantage of AAV-DJ (mouse only, NHP liver is not high) [10-27]; AAV-LK03 human hepatocyte tropism (only transduced humanized mouse\u002Fhuman hepatocytes, apparent transcriptional silencing of mouse hepatocytes) [10-20]; liver tropism of AAV6.2FF\u002FLK03 (only NHP effective, not mice) [10-27]; brain endothelial tropism of AAV9-X1 (mouse CNS endothelium is strong, NHP cell type distribution is different) [10-9]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Examples of cross-species decay. \u003C\u002Fstrong>Yang 2024 showed that although capsid DNA distribution was similar between mice and monkeys, RNA\u002Fprotein expression was significantly lower in 20 of 28 tissues in monkeys than in mice (liver 66.5%→0.33%, heart 54%→7.8%, muscle 79%→34%, midbrain 30%→6%, tdTomato⁺ cell ratio) [10-27]. That is, high transduction in mice ≠ high transduction in humans\u002Fmonkeys, and apparent transcriptional silencing is an important bottleneck in the low-dose monkey model. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Tissue distribution of commonly used NHP serotypes. \u003C\u002Fstrong>AAV8 IV is highest in the whole body liver, and the liver is still the largest peripheral pool after IT [10-45]; AAV9 NHP is high in liver, heart, and skeletal muscle after IV, ICM\u002FIT has extensive neuronal and glial transduction in the whole brain + spinal cord (Zolgensma clinical basis), neonatal\u002Ffetal NHP neuronal transduction is better than that in adults [10-45][10-46]; AAVrh.10 NHP Liver and muscle are high, and heart expression is lower than AAV9 in head-to-head barcode studies [10-36]; AAVrh74 NHP skeletal muscle is high (Elevidys clinical 2×10¹⁴ vg\u002Fkg grade), and liver is the main off-target [10-37][10-38]; MyoAAV-2A NHP muscle mRNA is 10-50 higher than the native type. times, liver off-target reduction [10-44]. \u003C\u002Fp>","2026-09-18 11:13:51","2026-10-19 12:56:36",{"id":60,"service_pic":7,"service_title":61,"service_class":41,"class_name":42,"service_content":62,"service_tag":44,"created_at":63,"updated_at":64},10,"Six-Variable Decision Framework","\u003Cp>\u003Cstrong>Tissues and Cells - Decision Logic. \u003C\u002Fstrong>Confirm the target organ first, and then confirm the target cell type within the organ; the tropism of hepatocytes vs Kupffer vs sinusoidal endothelium, neurons vs astrocytes vs endothelium in the same organ can be completely different. Process: First determine the target cells → check the cell receptor expression → select the matching serotype → verify cell level transduction. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. \u003C\u002Fstrong>Equate \"tissue vg copy number\" to \"target cell expression\". High vg may simply be that the capsid is retained by vascular endothelial cells\u002Fmacrophages and does not enter the target cell; it must be verified with single-cell resolution (scRNA-seq, spatial transcriptome, cell-specific reporter) and attention should be paid to whether AAVR expression is in place. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>Vg can be measured in the brain after IV administration of AAV9, but neuronal transduction in the adult brain parenchyma is limited, mainly in the endothelium\u002Fglia; PHP.eB really enters the neurons [10-21][10-25]. \u003C\u002Fp>\n\u003Ch3>Species and strains\u003C\u002Fh3>\n\u003Cp>\u003Cstrong>Decision logic. \u003C\u002Fstrong>First check whether the receptor of the target cell (Ly6A? TfR1? Galactose?) exists and is accessible in the species\u002Fstrain used; give priority to serotypes that are conserved among species (8\u002F9\u002Frh10\u002Frh74\u002FMyoAAV). Those that are not conserved must be verified in the target species. C57BL\u002F6J≠BALB\u002Fc≠NHP≠people. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. \u003C\u002Fstrong>Direct copy of mouse PHP.B to NHP\u002Fhuman. PHP.B is highly transduced in the whole brain of C57BL\u002F6J, and NHP disappears completely[10-40][10-27]; BI-hTFR1 is ineffective in ordinary mice and requires human TfR1 KI[10-43]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>Liver targeting selection AAV8\u002Frh10\u002Frh74 (conserved across species); do not default to PHP.B for BBB crossing. Effective in NHP. \u003C\u002Fp>\n\u003Ch3>Method of administration\u003C\u002Fh3>\n\u003Cp>\u003Cstrong>Decision logic. The\u003C\u002Fstrong>route determines receptor accessibility (luminal vs basolateral, vitreous vs subretinal). Process: Select the optimal path according to the target tissue → Confirm the serotype affinity under this path (may be different from IV) → Match verification. The same capsid IV affinity can be reversed with topical administration. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. IV and topical administration are thought to be equally effective. AAV2 IV is extremely low in the brain but highly effective when injected into the brain parenchyma; AAV6 is optimal for airway luminal administration, while IV is mainly for the liver and heart; AAV4 ICV is partial to the ependyma, and IV is partial to the lungs\u002Fkidneys\u002Fpancreatic islets. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>AAV9 IV administration mainly targets the liver and heart, while local injection into the myocardium directly enriches the myocardium; retinal SR→RPE\u002Fphotoreceptor, IVT→ganglion mainly requires penetrating types such as AAV2.7m8 to reach the outer layer [10-47]. \u003C\u002Fp>\n\u003Ch3>Dose and time\u003C\u002Fh3>\n\u003Cp>\u003Cstrong>Decision logic. \u003C\u002Fstrong>The affinity is more \"pure\" at low doses (before receptor saturation). At high doses, hepatic non-specific interception increases and the proportion of positive cells increases, but off-target and innate immune burden increase. The time window determines the peak expression (AAV1\u002F6\u002F7\u002F8\u002F9 is fast, AAV2\u002F3\u002F4\u002F5 is slow) [10-26]; the time window is different between neonates and adults [10-25]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. \u003C\u002Fstrong>Thought higher doses must be better. High-dose AAV9 IV can cause elevated liver transaminases and dorsal root ganglion toxicity (NHP\u002Fperson); be wary of dose-dependent tropism shifts. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>PHP.eB can reach 69% of cortical neurons at 1×10¹¹ vg, without blindly increasing the dosage [10-21]. \u003C\u002Fp>\n\u003Ch3>Promoter\u003C\u002Fh3>\n\u003Cp>\u003Cstrong>Decision logic. \u003C\u002Fstrong>The capsid determines \"whether it can enter the cell\", and the promoter determines \"where it is expressed after entering.\" The two work together: capsid enters muscle + MHCK7 = muscle specific; capsid enters broadly + synapsin\u002FGFAP = neuron\u002Fglia restricted. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. \u003C\u002Fstrong>It is believed that the promoter can completely make up for the lack of capsid tropism. If the capsid does not enter the target cell at all, no matter how strong the tissue-specific promoter is, there will be no substrate; on the contrary, if the capsid enters the liver non-specifically, the strong promoter will only amplify off-target expression. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>rh74+MHCK7 (Elevidys) achieves high muscle expression, but if the capsid does not effectively enter the muscle fiber, the promoter cannot save the day [10-37]. \u003C\u002Fp>\n\u003Ch3>Study endpoint\u003C\u002Fh3>\n\u003Cp>\u003Cstrong>Decision logic. \u003C\u002Fstrong>Select molecules (vg\u002FmRNA), cells, tissues, and functional multi-layered endpoints based on affinity and target cells, and match expression kinetics with sampling time. If the capsid is endothelial, the endpoint should measure vascular function rather than neuronal electrophysiology; if the capsid is endothelial, the endpoint should measure glial activation rather than behavioral improvement. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Common Misconceptions. \u003C\u002Fstrong>Only look at fluorescence intensity without looking at quantitative expression and function, and ignore non-target tissue sampling. GFP positivity ≠ effective therapeutic protein expression ≠ functional improvement; vg copy number, mRNA, protein, and function should be verified step by step. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Example. \u003C\u002Fstrong>In DMD studies, rh74 microdystrophin needs to simultaneously measure muscle histology, muscle strength and liver function (off-target toxicity), not just fluorescence [10-38]. \u003C\u002Fp>","2026-09-18 11:14:35","2026-10-16 21:49:34",{"id":66,"service_pic":7,"service_title":67,"service_class":41,"class_name":42,"service_content":68,"service_tag":44,"created_at":69,"updated_at":70},11,"How Engineered Capsids Alter Tropism","\u003Cp>\u003Cstrong>Receptor reprogramming. \u003C\u002Fstrong>The essence of receptor reprogramming is to \"replace receptors rather than cells.\" It usually retains AAVR dependence and adds a new target enrichment pathway. The PHP.B family inserts a 7-mer peptide into AAV9 VR-IV to obtain Ly6A binding, adding a \"trans-brain endothelial Ly6A transcellular\" pathway [10-22]; subsequent PHP.eB optimization efficiency [10-21], PHP.V1 partial to brain endothelium, PHP.N partial to neurons, PHP.A partial to glial and liver decreased by 152 times [10-9]. MyoAAV\u002FAAVMYO inserts the RGD motif, actively binds to integrin αVβ6\u002FαVβ8\u002Fα5β1, and reprograms AAV9 from galactose-dependent to integrin-dependent. Muscle mRNA is 10-50 times higher than AAV9 (NHP), liver off-target is reduced, and AAVR is still required [10-44][10-52]. BI-hTFR1 binds to the luminal epitope of human TfR1 to achieve trans-BBB receptor-mediated transcellular [10-43]. In terms of lung targeting, AAV-QPE\u002FVNT and others achieve lung redirection by destroying AAV2 HSPG binding, and AAV2.5T (2\u002F5 chimeric) is efficiently transferred to the airway epithelium through the airway lumen [10-9]; 9P31\u002F9P36 binds to primate conserved carbonic anhydrase IV (CA-IV) [10-9]. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Peptide insertion. \u003C\u002Fstrong>Inserting targeting peptides into VR-IV\u002FVR-VIII (GH loop) is the most commonly used method to change the affinity: RGD peptide → integrin binding (muscle\u002Ftumor blood vessels) [10-44]; TLAVPFK (PHP.B) → Ly6A [10-22]; AAV2.7m8 (AAP framework mutation + peptide display) → Intravitreal administration penetrates all layers of the retina, and phototransduction can reach about 60% (natural type only) 2-20%) [10-47]; DJ-NSS\u002FDJ-MVN intranasal administration redirects to alveolar epithelial\u002Falveolar macrophages respectively [10-9]. The inserted peptide must be exposed to the surface of the capsid tripod protrusions without disrupting AAVR binding and assembly. \u003C\u002Fp>\n\u003Cp>\u003Cstrong>Filter sources (brief description). \u003C\u002Fstrong>CREATES\u002FCre relies on selection to enrich the correct capsids in specific cell types of Cre mice (PHP.B source) [10-22]; DELIVER in vivo directed evolution to retain muscle tropism across species (MyoAAV) [10-44]; barcode in vivo screening 21-100+ capsid pools in one injection, barcode sequencing to quantify tissue DNA\u002FRNA (Yang 2024 paradigm) [10-27]; transcriptome-assisted screening (scRNA-seq\u002Fspatial transcriptome) analyzes single-cell level tropism [10-48]; humanized models (human liver chimeric mouse LK03 [10-20], human TfR1 KI mouse [10-43], human BBB chip) solve the problem of \"mouse screening is effective, but human screening is invalid\". See Chapter 12 for detailed screening procedures. \u003C\u002Fp>","2026-09-18 11:15:01","2026-10-17 23:18:46",{"id":72,"service_pic":7,"service_title":73,"service_class":41,"class_name":42,"service_content":74,"service_tag":44,"created_at":75,"updated_at":76},12,"Serotype Selection Workflow and Verification Checklist","\u003Cp>Follow the following 10 steps to check one by one. If any one is not satisfied, change to the next candidate. \u003C\u002Fp>\n\u003Col>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Clear the target cell type (not an organ, but a specific cell, such as hepatocyte\u002Fmotor neuron\u002Fphotoreceptor\u002FRPE). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Check the receptor\u002Fco-receptor expression of the cell in the target species (AAVR must have; attachment receptor? Ly6A? PDGFR? Galactose? TfR1?). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Check the evidence that the serotype already exists in this cell + this species + the proposed pathway, and give priority to Level A evidence. \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Preliminarily screen 2-3 candidate serotypes (find the organization + the pathway \"high\" in the 6.2 matrix, giving priority to 8\u002F9\u002Frh10\u002Frh74\u002FMyoAAV, which is conserved across species). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Verify the tropism based on the route of administration - the tropism of different routes for the same serotype may be different. \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Check that the promoter cooperates with the capsid (whether the promoter can be transcribed after the capsid enters the cell, do not expect the promoter to make up for the capsid). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Determine dose window (effective dose vs toxic dose, considering liver\u002FDRG off-target and immune burden). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Set study endpoints (vg→mRNA→protein→functional multilayer, single cell resolution, sampling time to match expression kinetics). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Assess risk of species extrapolation (distinguish conservative vs. non-conserved serotypes; key conclusions need to be reviewed in NHP or humanized models). \u003C\u002Fli>\n\u003Cli>\u003Cstrong> \u003C\u002Fstrong>Set controls (empty vector, non-target tissue sampling, head-to-head comparison of different serotypes; check the original text before using combinations marked \"to be verified\" in the matrix). \u003C\u002Fli>\n\u003C\u002Fol>","2026-09-18 11:15:21","2026-10-17 19:54:25",{"id":78,"service_pic":7,"service_title":79,"service_class":41,"class_name":42,"service_content":80,"service_tag":44,"created_at":81,"updated_at":82},13,"References for this chapter","\u003Cp>[10-1] Hordeaux J, et al. 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DOI:10.1038\u002Fs41596-018-0097-3. [F]\u003C\u002Fp>","2026-09-18 11:15:49","2026-10-23 16:58:04",{"id":84,"service_pic":7,"service_title":85,"service_class":6,"class_name":9,"service_content":86,"service_tag":11,"created_at":87,"updated_at":88},14,"Promoter and expression control selection","\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>\u003Cstrong>Type\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>\u003Cstrong>Applicable scenarios\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>\u003Cstrong>Verification required\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Broad spectrum promoter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>Maximize model coverage or rapid verification\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>Different tissue strengths, silencing, immunity and toxicity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Tissue\u002Fcell type promoter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>Restrict transcription to target cells\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>Species conservation, size and activity in disease states\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Enhancer + minimal promoter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>More fine cell subtype selection\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>Orientation\u002Fcopy number, cross-species migration and leakage\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>miRNA de-targeting\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>Depress non-target expression in liver, immune cells, etc.\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>Endogenous miRNA abundance, saturation and tissue differences\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"115\">\n\u003Cp>Induction system\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"226\">\n\u003Cp>Time control or reversible adjustment\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"254\">\n\u003Cp>Leaky expression, inducer exposure and expression cassette capacity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>Cross-species enhancer research from 2016 to 2020 proved that short regulatory elements can achieve finer neuronal subtype selection than traditional large promoters; however, enhancer performance is still affected by species, brain regions, load and minimal promoter combinations. Specificity, positive rate, expression intensity per unit cell and expression cassette size should be compared at the same time when selecting. [11-1 to 11-3]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028036\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[11-1] Dimidschstein J, et al. A viral strategy for targeting and manipulating interneurons across vertebrate species. Nat Neurosci. 2016;19:1743-1749. DOI:10.1038\u002Fnn.4430. PMID:27798629. [H]\u003C\u002Fp>\n\u003Cp>[11-2] Jüttner J, et al. Targeting neuronal and glial cell types with synthetic promoter AAVs in mice, non-human primates and humans. Nat Neurosci. 2019;22:1345-1356. DOI:10.1038\u002Fs41593-019-0431-2. PMID:31285614. [H]\u003C\u002Fp>\n\u003Cp>[11-3] Vormstein-Schneider D, et al. Viral manipulation of functionally distinct interneurons in mice, non-human primates and humans. Nat Neurosci. 2020;23:1629-1636. DOI:10.1038\u002Fs41593-020-0692-9. PMID:32807948. [H]\u003C\u002Fp>\n\u003Cp>[11-4] Wang JH, et al. AAV as a delivery vector for gene therapy of human diseases. Signal Transduct Target Ther. 2024;9:78. DOI:10.1038\u002Fs41392-024-01780-w. [N\u002FH]\u003C\u002Fp>","2026-09-18 11:16:14","2026-10-28 23:08:54",{"id":90,"service_pic":7,"service_title":91,"service_class":92,"class_name":93,"service_content":94,"service_tag":95,"created_at":96,"updated_at":97},16,"Quick guide to route of administration",15,"Administration Routes, Dosing, and Expression Assessment","\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>\u003Cstrong>Path\u002FPart\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>\u003Cstrong>Mouse dosage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>\u003Cstrong>Rat dosage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>\u003Cstrong>NHP\u002FLarge Animal Dosage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>\u003Cstrong>Common volumes (mice)\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>\u003Cstrong>First expression observation\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Tail vein (IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-1×10^12 vg\u002Fpiece (about 5×10^12-5×10^13 vg\u002Fkg)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>CNS targeting AAV9 3×10^13-1×10^14 vg\u002Fkg\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^12-6×10^13 vg\u002Fkg (system IV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>100-300 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Retroorbital vein (RO)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-2×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>Generally not recommended\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>—\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>100-150 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Portal vein (PV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^10-1×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^11-1×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>~10^12-10^13 vg\u002Fkg (historical plan, to be verified)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>≤100 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intraperitoneal cavity (IP)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-1×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>2×10^11 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>—\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>100-200 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Subcutaneous (SC)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>5×10^10-4×10^11 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>To be verified (approximately 1×10^12 vg\u002Fkg level)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>~10^12 vg level\u002Fpart (to be verified)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>~100 μL\u002Fdot\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>≥6 months\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intramuscular (IM)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^9-1×10^11 vg\u002Fmuscle; total multiple muscles 1×10^11-1×10^12\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>~1×10^11 vg\u002Fmuscle\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>rhesus IM AAV8 3×10^12 GC\u002Fkg\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>25-50 μL\u002Fdot\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Brain stereotaxy\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^9-1×10^10 vg\u002Fsite\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>2×10^10-6×10^10 vg\u002Fsite\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1.5×10^11 vg\u002Fbrain (putamen)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>0.2-2 μL\u002Fsite\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Ventricles (neonatal\u002Fadult ICV)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Newborn 1-2×10^10 GC\u002Fhemisphere; adult 1×10^11 vg\u002Fside\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>Stereotaxy of adult lateral ventricles\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>4×10^13 vg (bilateral)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>Newborn 1-2 μL\u002Fside; adult 5-10 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intrathecal (IT\u002Flumbar puncture)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>5×10^10-1×10^11 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1-2×10^11 vg\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>4.8×10^12-9.2×10^13 vg\u002Fpiece; same level as dog\u002Fpig\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>5-10 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Cerebellomedullary cistern (CM\u002FICM)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^10-7×10^10 vg\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^12 vg\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1.7-5×10^13 GC\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>1-10 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Lumbar puncture (LP)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Same as IT(L5-L6)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>Same as IT\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^13 vg\u002Fkg; pig 1.4×10^13 GC\u002FmL×400-800 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>5-10 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Subretinal (SR)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^8-3×10^9 vg\u002Feye\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^9-1×10^10 vg\u002Feye\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>7×10^11-7×10^12 vg\u002Feye (AAV8); dog 1-1.4×10^11\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>1-2 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intravitreal cavity (IVT)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^9-1×10^10 vg\u002Feye\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^9-5×10^9 vg\u002Feye\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>7×10^11 vg\u002Feye\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>1-2 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intranasal\u002FIntratracheal\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>4×10^11-1×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^11-5×10^12 vg\u002Fonly\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>NHP atomization ~3 mL; sheep 10 mL\u002Ftime\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>25-50 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>1-2 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Renal vein\u002Fretrograde ureter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-1×10^12 vg\u002Fkidney\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^11-5×10^11 vg\u002Fkidney\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Data is limited\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>100 μL\u002Fkidney\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Pancreatic duct\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>2×10^11-2×10^12 vg\u002Fonly (AAV6)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Limited data (ERCP path)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>100 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>4-6 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Articular cavity (IA)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>0.5×10^10-1×10^11 vg\u002Fjoint\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^10-2.5×10^10 vg\u002Fjoint\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Clinical magnitude of equine OA\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>5 μL\u002Fjoint\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intratumoral (ITu)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>5×10^10-1×10^11 vg\u002Ftumor\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^10-5×10^10 vg\u002Ftumor\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Spontaneous tumor in dogs (to be diagnosed)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>50-100 μL\u002Ftumor\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>1-2 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Sciatic nerve\u002Fperipheral nerve\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>2.1×10^8 vg(scAAV1)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>2×10^11 GC\u002Fnerve\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>2.5×10^12-2.6×10^13 vg\u002Fnerve\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>2-20 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Cochlea\u002Finner ear\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^10-5×10^10 vg\u002Fear\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>About 2-5 μL (to be nucleated)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-7×10^11 vg\u002Fear\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>0.6-2 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Myocardial part\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>5×10^10-1×10^11 vg\u002Fheart\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>1×10^11-5×10^11 vg\u002Fheart\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Canine 7-Fr endocardial catheter; pig multi-site thoracotomy\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>30 μL(3 points)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Fat pad\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>1×10^11-2×10^11 vg\u002Fpad\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>To be approved\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>—\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>30-50 μL\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>3-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"113\">\n\u003Cp>Intracutaneous\u002Fwound\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>~2×10^10 vg\u002Fdot\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"107\">\n\u003Cp>50-100 μL per point\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"127\">\n\u003Cp>Pig skin transformation model\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"93\">\n\u003Cp>10-20 μL\u002Fdot\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"73\">\n\u003Cp>2-4 weeks\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>","AAV, Administration Routes, Dosing, and Expression Assessment","2026-09-18 11:17:10","2026-10-23 19:54:47",{"id":99,"service_pic":7,"service_title":100,"service_class":101,"class_name":102,"service_content":103,"service_tag":104,"created_at":105,"updated_at":106},18,"IV tail vein",17,"Detailed Administration-Route Parameters and Coverage","\u003Cp>Applicable species and strains: mice C57BL\u002F6J, BALB\u002Fc, CD-1 (6-8 weeks, 18-25 g); rats Sprague-Dawley, Wistar (6-8 weeks, 150-250 g) [12-1,12-8]. Dose range: mouse liver\u002Fheart\u002Fmuscle targeting 1×10^11-1×10^12 vg\u002Fmouse (approximately 5×10^12-5×10^13 vg\u002Fkg), BBB-penetrating engineering capsid (PHP.B type) 1×10^11-1×10^12 vg\u002Fmouse; rat CNS targeting AAV9 3×10^13-1×10^14 vg\u002Fkg[12-1,12-7,12-8]. Injection volume: 100-300 μL for mice (the upper limit is about 10 mL\u002Fkg); 200-500 μL for rats [12-1]. Needles: 27-30 G insulin syringe for mice; 28-30 G catheter or 25 G needle for rats [12-1]. Speed: manual slow injection for 30-60 s, fast bolus injection is prohibited [12-1]. Anesthesia: Awake operation with heated fixator; PHP.B protocol polyisoflurane inhalation anesthesia [12-1]. Operating points: Expand the tail vein in a 37°C water bath before injection, insert the needle from the lateral\u002Fdorsal vein of the distal 1\u002F3 of the tail (\u003C15°), the bolus is successful if there is no resistance and the vein turns white, and the cotton ball is pressed for 10-30 s; the interval between multiple injections is ≥ 2 weeks, and repeated administration is limited by anti-capsid neutralizing antibodies [12-1, 12-12]. \u003C\u002Fp>\n\u003Cp>Coverage: liver (AAV8\u002F9 is the strongest, 90%+ hepatocytes with dose), heart (AAV9 preferentially myocardium), skeletal muscle; AAV9 and engineered capsids can cross the BBB; expression peaks in 2-6 weeks and lasts for a long time [12-6]. Main off-target: liver is the largest off-target organ, followed by spleen, kidney, and lung; high-dose DRG neurons are enriched [12-11]. Limitations: Strong dependence on mouse strains (AAV-PHP.B is only effective in C57BL\u002F6J lines and related to Ly6a) [12-7]; see section 8.4 for the risk of high-dose hepatotoxicity\u002Fcomplement activation. \u003C\u002Fp>","AAV, Detailed Administration-Route Parameters and Coverage","2026-09-18 11:19:13","2026-10-16 21:31:45",{"id":108,"service_pic":7,"service_title":109,"service_class":101,"class_name":102,"service_content":110,"service_tag":104,"created_at":111,"updated_at":112},19,"Retro-orbital IV","\u003Cp>Applicable species: only mice (adults and newborns), rats are generally not recommended (high risk of orbital anatomy) [12-9]. Dose range: equivalent to tail vein IV, commonly used is 1×10^11-2×10^12 vg\u002Fanimal; literature on neonatal rat AAV9 uses 1.5×10^14 vg\u002Fkg[12-10]. Injection volume: 100-150 μL\u002Feye for adult mice (the upper limit for a single injection is 200 μL or 1% of body weight, whichever is smaller); neonatal mice ≤ 10 μL [12-9]. Needle: 27-32 G (29-30 G is commonly used) [12-9]. Speed: Inject slowly for 10-30 s, wait for 2-3 s before withdrawing the needle [12-9]. Anesthesia: General anesthesia is required (isoflurane inhalation is preferred) [12-9]. Operational points: After anesthesia, the head is tilted towards the operator, and the neck is gently pressed to make the retroorbital venous plexus congested with blood. The needle is inserted from the medial canthus along the orbital wall at 45° to the dorsal side for 2-3 mm, and blood is sucked back to confirm the insertion of the plexus; the same eye should be used ≤ 2 times, and the two eyes should be alternated with an interval of ≥ 2 days [12-9]. Coverage: equivalent to system IV (liver, heart, skeletal muscle, BBB); off-target includes occasional inflammation of periocular tissues and Harderian glands. Limitations: There is a risk of eyeball damage, retroorbital hematoma, and keratitis, and some institutions have restricted non-essential use [12-9]. \u003C\u002Fp>","2026-09-18 11:19:41","2026-10-18 02:55:19",{"id":114,"service_pic":7,"service_title":115,"service_class":101,"class_name":102,"service_content":116,"service_tag":104,"created_at":117,"updated_at":118},20,"Portal vein injection","\u003Cp>Applicable species: mice, rats; NHP is a historical preclinical\u002Fclinical protocol (has been basically replaced by peripheral IV) [12-13]. Dose range: 1×10^10-1×10^12 vg\u002Fmouse (AAV8 can transduce 90-95% of hepatocytes through the portal vein); 1×10^11-1×10^12 vg\u002Fmouse [12-13]. Injection volume: direct portal vein puncture ≤100 μL; rat 100-200 μL[12-13]. Needle: mouse 1 mL syringe with 33 G needle, or transsplenic 32 G needle [12-13,12-14]. Speed: Slowly inject for 1-2 minutes [12-14]. Anesthesia: Isoflurane inhalation + laparotomy, aseptic operation, and postoperative analgesia are required [12-13]. Operational points: Make a midline incision of 1-1.5 cm in the abdomen, identify the portal vein (the hepatic portal is accompanied by the bile duct), insert a 33 G needle above the confluence of the gastroduodenum, and slowly inject ≤100 μL. After the needle is withdrawn, press with a sterile cotton swab for 3-5 minutes to stop bleeding; the minimally invasive alternative is to inject 50 μL slowly through the spleen parenchyma (lower risk of bleeding) [12-14]. Coverage: hepatocytes (AAV8\u002F9 up to 90%+), liver non-parenchymal cells are also transduced, liver lobule distribution is zonal (opposite to NHP in mice) [12-15]; much lower than systemic off-target of peripheral IV. Limitations: Laparotomy mortality and recovery costs, risk of portal vein bleeding\u002Fthrombosis, non-repetitive administration [12-13]. \u003C\u002Fp>","2026-09-18 11:20:04","2026-10-19 13:35:32",{"id":120,"service_pic":7,"service_title":121,"service_class":101,"class_name":102,"service_content":122,"service_tag":104,"created_at":123,"updated_at":124},21,"Intraperitoneal (IP)","\u003Cp>Applicable species: mice, rats [12-16,12-17]. Dose range: 1×10^11-1×10^12 vg\u002Fmouse; 2×10^11 vg\u002Fmouse [12-16,12-17]. Injection volume: mice 5-10 mL\u002Fkg (approximately 100-200 μL); rats 10-20 mL\u002Fkg (200 μL in literature) [12-16,12-17]. Needle: mouse 25-27 G; rat 23-25 ​​G[12-16]. Speed: Just give it a quick bolus in a few seconds [12-16]. Anesthesia: conscious or light sedation, good immobilization is required [12-16]. Key points of the operation: Fix the head in a low position to move the organs upward, insert a 25 G needle into the left lower abdomen (avoiding the bladder and liver), withdraw intestinal fluid\u002Furine\u002Fblood after feeling empty, and then inject again; it can be used to target the liver of neonatal\u002Fyoung mice [12-16]. Coverage: It absorbs into the blood through diaphragm lymph\u002Fperitoneal capillaries and then enters the liver through the portal vein. The liver transduction is lower than IV, but the diaphragm, fat, and pancreas transduction is enhanced (AAV6.2, AAV-DJ is obvious), and the rat colon mucosa\u002Fsubmucosa can also be transduced [12-16,12-17]; the peak expression is 2-6 weeks. Limitations: Bioavailability is unstable (affected by abdominal contents, body position, inflammation), and is not suitable for CNS projects that require precise dosage [12-16]. \u003C\u002Fp>","2026-09-18 11:20:23","2026-10-19 10:49:54",{"id":126,"service_pic":7,"service_title":127,"service_class":101,"class_name":102,"service_content":128,"service_tag":104,"created_at":129,"updated_at":130},22,"Subcutaneous injection (Subcutaneous, SC)","\u003Cp>Applicable species: mice; rat\u002FNHP data to be verified[12-18]. Dose range: 5×10^10-4×10^11 vg\u002Fmouse (fat pad local protocol 2×10^10 vg\u002Fside) [12-18]. Injection volume: approximately 100 μL\u002Fpoint; 25-30 μL\u002Fpoint in local fat pad [12-18]. Needle: 30-31 G (3\u002F10 mL syringe, needle length 8 mm) [12-18]. Speed: Slowly inject for 10-30 s to form skin cells [12-18]. Anesthesia: Just stay awake and immobilized. Coverage: subcutaneous loose connective tissue, local fat (iWAT\u002FiBAT); liver expression after absorption into the bloodstream - although the bioavailability of SC is \u003C100%, the specific activity of liver vg is higher than portal injection, which can be compensated; the expression can last for ≥ 6 months [12-18]. Limitations: slow absorption and large variability, not suitable for experiments that require rapid peaking; local inflammation\u002Finduration affects film reading; not recommended as the first choice for systemic liver targeting [12-18]. \u003C\u002Fp>","2026-09-18 11:20:41","2026-10-16 21:56:10",{"id":132,"service_pic":7,"service_title":133,"service_class":101,"class_name":102,"service_content":134,"service_tag":104,"created_at":135,"updated_at":135},23,"Intramuscular (IM)","\u003Cp>Applicable species: mice, rats, NHP, dogs [12-19,12-20,12-23]. Dose range: mouse single point 1×10^9-1×10^11 vg\u002Fmuscle, multi-muscle strategy total 1×10^11-1×10^12 vg\u002Fmouse; rat single point approximately 1×10^11 vg\u002Fmuscle; rhesus IM AAV8 3×10^12 GC\u002Fkg divided into bilateral vastus lateralis muscles [12-19,12-20]. Injection volume: 25-50 μL for a single point in mice; 0.5-1 mL\u002Fpoint for NHP, and the total volume can reach 4-135 mL[12-20,12-22]. Needles: Mouse 29-30 G Hamilton; NHP 22-25 G long needle [12-20]. Speed: Inject slowly for 30-60 s, and wait for 10-20 s after withdrawing the needle to prevent reflux [12-19]. Anesthesia: light anesthesia (isoflurane\u002FAvertin); NHP sedation\u002Fgeneral anesthesia. Operation points: After hair removal and disinfection of mice, insert a needle 3-5 mm along the direction of muscle fibers; NHP has multiple points on both quadriceps\u002Fgluteal muscles with a spacing of ≥1 cm, and the injection points are marked with ink [12-21,12-22]. Coverage: Injected muscle transduction (mainly muscle fibers), distal muscles are not transduced; secreted transgenes can enter the bloodstream; NHP IM AAV8 has unexpectedly significant liver targeting (the number of liver GCs is equivalent to that of local muscle) [12-20]; the peak expression is 2-6 weeks, and loss of NHP immune-mediated expression is common but can be restored [12-24]. Multi-muscle strategy: Multiple scattered injections cover more muscle fibers, and the upper limit of the total dose is limited by liver toxicity; regional limb perfusion (VLP\u002FIAPD) can be used as an alternative for large animals [12-22,12-23]. \u003C\u002Fp>","2026-09-18 11:20:59",{"id":137,"service_pic":7,"service_title":138,"service_class":101,"class_name":102,"service_content":139,"service_tag":104,"created_at":140,"updated_at":141},24,"Stereotaxic brain injection (Intracerebral\u002FIntraparenchymal)","\u003Cp>Applicable species: mice (C57BL\u002F6, FVB, etc.), rats (SD, Long-Evans), NHP (rhesus\u002Fcynomolgus) [12-1]. Dose range: adult mouse striatum 1×10^9-5×10^10 vg\u002Fsite, hippocampus 1×10^9-1×10^10 vg\u002Fside; rat striatum 2-6×10^10 vg\u002Fsite; NHP putamen 1.5×10^11 vg\u002Fbrain (2×50 μL) [12-1]. Injection volume: mouse 0.2-2 μL\u002Fsite (cumulative ≤5 μL\u002Fbrain); rat 0.5-2 μL; NHP 50-150 μL\u002Fsite[12-1]. Needle: Mouse Hamilton 7000 series equipped with 33 G stainless steel needle or glass microelectrode; NHP customized fused-silica anti-reflux catheter [12-1]. Speed: 100-200 nL\u002Fmin is most commonly used for mice; NHP ramp-up 0.2→0.5→1.0→3.0 μL\u002Fmin[12-1,12-40]. Anesthesia: Mice were treated with isoflurane 3-5% for induction and 1-2% maintenance, intraoperative thermostat and postoperative buprenorphine analgesia; NHP isoflurane 1-3% for tracheal intubation monitoring [12-1]. Key points of operation: The height difference between bregma and lambda is \u003C0.05 mm, drill and thin the skull to open the dura mater, leave the needle for 5-10 minutes after injection and then withdraw the needle at 1 mm\u002Fmin to reduce reflux [12-1]. Coverage: Diffusion radius is about 1-2 mm (half-life distance 500-1000 μm), limited to around the injection site + anterograde\u002Fretrograde projection pathway; cell type is determined by the promoter; peak expression is 2-4 weeks [12-1]. Safety: needle tract bleeding, high-dose epilepsy in cortex\u002Fhippocampus; direct neurotoxicity has been reported in NHP intracerebral injection of AAVrh8 [12-38]. \u003C\u002Fp>","2026-09-18 11:21:19","2026-10-19 12:35:35",{"id":143,"service_pic":7,"service_title":144,"service_class":101,"class_name":102,"service_content":145,"service_tag":104,"created_at":146,"updated_at":147},25,"Intracerebroventricular (ICV)","\u003Cp>Applicable species: newborn mice P0-P4, adult mice\u002Frats, NHP[12-33,12-39]. Dose range: neonatal P0-P2 1-2×10^10 GC\u002Fhemisphere (total 2-4×10^10\u002Fmouse); adult mice 1×10^11 vg\u002Fside; NHP 4×10^13 vg (bilateral, 2×10^13\u002Fventricular×1 mL) [12-33]. Injection volume: neonates 1-2 μL\u002Fside (P0 recommendation ≤1 μL\u002Fside); adults 5-10 μL\u002Fside; NHP 1 mL\u002Fventricular [12-33]. Needles: newborn 10 μL Hamilton with 32 G needle; adult Kopf stereotaxic + Hamilton 33 G bevel needle; NHP CT\u002Ffluoroscopic guided lumbar puncture needle\u002Fventricular catheter [12-33]. Speed: newborn ≤30 nL\u002Fs (no exceeding 15 nL\u002Fs); adults 1 μL\u002Fmin[12-33]. Anesthesia: neonatal hypothermic anesthesia on ice for 60 s; adult isoflurane + stereotaxic frame; NHP isoflurane [12-33]. Key points of operation: Percutaneous translucent positioning of neonates (lambda suture to 2\u002F5 of the eye, opening 0.25 mm and inserting needle 2 mm); drilling holes in adults, selecting the dural needle and retaining the needle for 2 minutes; P0 injection of neonates has the widest neuron transduction, and the proportion of astrocytes increases after P3 [12-33]. Coverage: neonates extensively cover the cortex, hippocampus, striatum, brainstem, cerebellum, spinal cord lumbar segment > thoracic segment > cervical segment; in adults only the ependyma + choroid plexus + adjacent brain parenchyma; peak expression is 2-4 weeks [12-33]. Safety: Neonatal injection is too fast (>15 nL\u002Fs) or excessive (>2 μL\u002Fside) → increased intracranial pressure, cerebral hemorrhage, and 1-week mortality [12-33]. \u003C\u002Fp>","2026-09-18 11:21:40","2026-10-19 13:50:31",{"id":149,"service_pic":7,"service_title":150,"service_class":101,"class_name":102,"service_content":151,"service_tag":104,"created_at":152,"updated_at":153},26,"Intrathecal injection (Intrathecal, IT\u002Flumbar spine)","\u003Cp>Applicable species: mice, rats, NHP, dogs, pigs[12-32,12-34,12-35]. Dose range: adult mice 5×10^10 vg\u002Fmouse (bolus 5 μL), up to 3.35×10^11 vg\u002Fmouse; rat lumbar puncture bolus injection 100-200 μL; NHP IT range 4.8×10^12-9.2×10^13 vg\u002Fmouse (1.0-2.5 mL); pig 7×10^12 GC\u002Fonly[12-32,12-35]. Injection volume: mice 5-10 μL (standard 5 μL); NHP 1.0-2.5 mL; pigs 400-800 μL\u002Fsegment [12-32]. Needle: mouse 30 G min with iohexol contrast agent CT for confirmation [12-31,12-32]. Anesthesia: mice are awake and fixed (Hylden-Wilcox modification) or mild isoflurane; NHP isoflurane 1-3% lateral flexion position [12-32] Operation points: mice L5-L6 intervertebral space (located along the iliac crest line), sudden bounce on the tail = entering the subarachnoid space; NHP CSF reflux + contrast agent double confirmation, Trendelenburg head down 5-10 after bolus injection min can significantly improve brain\u002Fcervical segment coverage [12-30,12-32]: The coverage is the thickest in the lumbar segment, followed by the thoracic segment, and the weakest in the cervical segment; AAV9 transduces motor neurons + DRG sensory neurons + oligodendrocytes; the peak expression is 3-6 weeks [1 2-31]. Safety: Degeneration of DRG sensory neurons (most consistent target toxicity, see Section 8.4); excessive\u002Ftoo rapid → acute neurological deficit; fatal meningitis has occurred with pig AAV8-IL-10 [12-26,12-34].","2026-09-18 11:21:58","2026-10-21 16:40:31",{"id":155,"service_pic":7,"service_title":156,"service_class":101,"class_name":102,"service_content":157,"service_tag":104,"created_at":158,"updated_at":159},27,"Cisterna magna injection (Cisterna Magna, CM\u002FICM)","\u003Cp>Applicable species: newborn\u002Fadult mice, rats, NHP, cats\u002Fdogs\u002Fpig\u002Fsheep[12-29,12-36]. Dose range: neonatal P1 mice 1×10^10 vg; adult mice 1×10^10-7×10^10 vg; rats 1×10^12 vg; NHP rhesus GLP low dose 1.7×10^13 GC, high dose 5×10^13 GC[12-29,12-36]. Injection volume: mice 1-10 μL; rats 50-100 μL; NHP 1-2 mL[12-29,12-36]. Needle: Mouse 33 G with Hamilton+ peristaltic pump; NHP 21-27 G Quincke lumbar puncture needle, C-arm fluoroscopy\u002FCT guidance [12-36]. Speed: 1 μL\u002Fmin for mice; 10 μL\u002Fmin for rats (100 μL CSF reserve volume is drawn first); NHP can push 1 mL within 30±5 s (approximately 2 mL\u002Fmin) [12-29,12-36]. Anesthesia: neonatal P1 5% isoflurane induction and 2% maintenance; rat ketamine + xylazine + acepromazine IP; NHP isoflurane lateral head flexion [12-29]. Operational points: The key risk is brainstem puncture (needle tip is too low\u002Ftoo deep → respiratory and cardiac arrest), and large animals must be guided by images; in rats, the needle hole is sealed with fibrinogen + thrombin; after surgery, Trendelenburg is inverted to increase brain coverage [12-30,12-36]. Coverage: ICM brain coverage is significantly wider than that of lumbar puncture LP - extensive cortex, hippocampus, cerebellum, brainstem; cervical\u002Fthoracic > lumbar segment (opposite to LP); high transduction of meningeal lymphatic vessels; peak expression at 3-4 weeks (scAAV9) [12-30,12-37]. Safety: brainstem puncture (most serious immediate risk), CSF leakage, DRG lesions, high-dose cerebellar toxicity, CSF lymphocytosis [12-36,12-38]. \u003C\u002Fp>","2026-09-18 11:22:16","2026-10-26 23:20:15",{"id":161,"service_pic":7,"service_title":162,"service_class":101,"class_name":102,"service_content":163,"service_tag":104,"created_at":164,"updated_at":165},28,"Lumbar Puncture (LP, large animals)","\u003Cp>Note: Mouse\u002Frat LP is the same operation as 8.2.9 IT (L5-L6 enters the subarachnoid space); this section focuses on the differences in large animal LP. Applicable species: NHP (cynomolgus\u002Frhesus), dogs, pigs [12-32]. Dose range: NHP juveniles 4.5×10^12 and 1.35×10^13 GC (2.5 mL); scAAV9-GFP 1×10^13 vg\u002Fkg; GLP 1.0\u002F3.0×10^13 vg\u002Fanimal [12-32]. Injection volume: NHP 2.5 mL standard (1-2 mL for adults); 1-5 mL for dogs; 400-800 μL\u002Fsegment for pigs (multiple points on the neck, chest, and waist are required) [12-32]. Needle: NHP 22 G spinal anesthesia catheter needle L4-L5 or L5-L6, CSF reflux + CT\u002Fiohexol double confirmation; dog\u002Fporcine 22 G spinal cord needle fluoroscopic guidance [12-32]. Speed: NHP is injected slowly for 1-2 minutes; postural effect: Trendelenburg can increase the motor neuron transduction rate to 73% → 80% in the waist and 29% → 55% in the neck 5-10 minutes after the injection [12-30,12-32]. Coverage: NHP LP spinal cord lumbar segment>thoracic segment>cervical segment, the brainstem\u002Fcerebellum is visible, and the cerebral cortex is weaker than ICM; single point coverage of the lumbar segment of pigs is insufficient and multiple points are needed [12-34]. Safety: See 8.4 DRG toxicity; postoperative headache, CSF leakage, and nerve root damage [12-26]. \u003C\u002Fp>","2026-09-18 11:22:36","2026-10-16 21:46:49",{"id":167,"service_pic":7,"service_title":168,"service_class":101,"class_name":102,"service_content":169,"service_tag":104,"created_at":170,"updated_at":171},29,"Subretinal injection (Subretinal, SR)","\u003Cp>Applicable species: mice, rats, NHP (rhesus\u002Fcynomolgus), dogs, pigs [12-45, 12-62, 12-44]. Dose range: mouse 1×10^8-3×10^9 vg\u002Feye; rat 1×10^9-1×10^10; NHP 7×10^11-7×10^12 vg\u002Feye (AAV8, engineered capsid can be as low as 1×10^10); dog 1.0-1.4×10^11 vg\u002Feye [12-45,12-62]. Injection volume: mice 1-2 μL; rats 2-5 μL; NHP 100-200 μL\u002Feye (traditional transretinal approach); dogs 200 μL\u002Feye [12-45,12-62]. Needle: Mouse 33 G Hamilton blunt needle or glass microelectrode; NHP 39 G injection cannula\u002F36-42 G retinal needle or 700 μm 30 G transscleral microneedle [12-5]. Speed: Inject bleb slowly, leaving the needle for 30 s after injection to prevent leakage; use MicroDose syringe pump for NHP [12-5]. Anesthesia: mouse isoflurane + preoperative mydriasis (1% tropicamide + 2.5% phenylephrine); NHP ketamine induction + isoflurane maintenance, preoperative dexamethasone + cefazolin [12-5]. Key points of the operation: The core is to create a controllable localized retinal detachment (bleb). AAV and Healon are mixed at a ratio of 1:1 to maintain the bleb shape; a 1 mm scleral pre-incision behind the limbus of the mouse; NHP standard three-incision vitrectomy through pars plana [12-5]. Coverage: RPE + photoreceptors in the bleb area (AAV8\u002F5 is the most efficient for photoreceptors), mouse bleb covers about 20-40% of the retina, and NHP single bleb covers about 20 μL around the fovea; the expression peaks at 3-6 weeks and lasts for >1.5 years [12-5]. Off-target: Bleb external transduction is extremely low, and leakage into the vitreous cavity causes RGC transduction. \u003C\u002Fp>","2026-09-18 11:22:58","2026-10-18 10:53:33",{"id":173,"service_pic":7,"service_title":174,"service_class":101,"class_name":102,"service_content":175,"service_tag":104,"created_at":176,"updated_at":177},30,"Intravitreal (IVT)","\u003Cp>Applicable species: mice, rats, NHP, dogs [12-47]. Dose range: mice 1×10^9-1×10^10 vg\u002Feye; rats 1×10^9-5×10^9; NHP 7×10^11 vg\u002Feye (AAV8) [12-47]. Injection volume: 1-2 μL for mice; 5 μL for rats; NHP 100 μL\u002Feye [12-47]. Needle: mouse 33 G Hamilton blunt needle; NHP 30 G μL allows diffusion; NHP punctures the anterior chamber and releases water to maintain intraocular pressure [12-47]. Coverage: Mainly transduces RGCs and part of the inner retina, AAV2 is the classic choice, and engineering capsids such as AAV2.7m8 can penetrate the ILM to transduce photoreceptors; the peak expression is 3-6 weeks [12-47]. Limitations: The internal limiting membrane (ILM) is the main physical barrier for IVT to transduce the outer retina; NHP vitreous body is about 2.5-4 mL, 0.1 mL is significantly diluted and requires a high-titer carrier; it is more likely to trigger systemic neutralizing antibody responses than SR; ILM ablation can significantly improve IVT efficiency [12-47].","2026-09-18 11:23:18","2026-10-29 01:02:44",{"id":179,"service_pic":7,"service_title":180,"service_class":101,"class_name":102,"service_content":181,"service_tag":104,"created_at":182,"updated_at":183},31,"Intranasal\u002FIntratracheal\u002FAerosol (Intranasal\u002FIntratracheal\u002FAerosol)","\u003Cp>Applicable species: mice, rats, NHP, sheep[12-48,12-49]. Dose range: mice 4×10^11-1×10^12 vg\u002Fmouse; rats 1×10^11-5×10^12; NHP reference clinical CFTR test 10^11-10^12 vg\u002Fkg (to be verified) [12-48,12-49]. Injection volume: mice i.t. 25-50 μL, i.n. 25-50 μL divided into both nostrils; rats 100-300 μL; NHP atomization approximately 3 mL\u002Fanimal (Aerogen Solo); sheep 10 mL\u002Ftime [12-48,12-49]. Needles: mouse video laryngoscope + Penn-Century micro-nebulizer, or 31 G tracheostomy needle \u002F 20 G transoral catheter; rat 22 G bulbous gavage needle [12-48,12-49]. Anesthesia: Mild inhalation of isoflurane (non-invasive); use Zoletil for invasive tracheotomy [12-48,12-49]. Key points of operation: non-invasive i.t. in supine position, place the tip of the nebulizer above the glottis through laryngoscope and inject; i.n. in the supine position, instill into the nostril and hold for 1 minute; instillation vs atomization: the infusion is unevenly distributed (more deposition in the lower lobe of the right lung) and the operation is fast, and the atomization distribution is even and close to physiology but the efficiency is only 10-30% [12-48,12-49]. Coverage: i.t. mainly transduces AT2, AT1 and bronchial epithelium, i.n. additionally transduces the upper respiratory tract; high doses (>2×10^12 vg) can penetrate the alveolar-capillary barrier into the blood and cause liver expression; the expression peaks in 1-2 weeks [12-48,12-49]. \u003C\u002Fp>","2026-09-18 11:24:08","2026-10-16 21:42:13",{"id":185,"service_pic":7,"service_title":186,"service_class":101,"class_name":102,"service_content":187,"service_tag":104,"created_at":188,"updated_at":189},32,"Renal Vein\u002FRetrograde Ureter (Renal Vein\u002FRetrograde Ureter)","\u003Cp>Applicable species: mice, rats; NHP\u002Fpig data are limited [12-50,12-51]. Dose range: retrograde ureter 1×10^11 vg\u002Fkidney; renal vein 1×10^11-1×10^12 vg\u002Fkidney (about half the tail vein dose); rat 1×10^11-5×10^11 vg\u002Fkidney [12-50,12-51]. Injection volume: approximately 100 μL\u002Fkidney; rat 100-200 μL\u002Fkidney [12-50,12-51]. Needle: tuberculin syringe + 33 G needle (retrograde ureter, fixed with 6-0 silk thread); 30-31 G needle for renal vein [12-50,12-51]. Speed: Slow retrograde ureteral injection of approximately 50-100 μL\u002Fmin [12-50,12-51]. Anesthesia: sodium pentobarbital 50 mg\u002Fkg IP, 37°C thermostat [12-50,12-51]. Key points of operation: Retrograde ureter requires a microvascular clamp to clamp the distal ureter and renal artery, first aspirate urine and then slowly inject it retrogradely into the renal pelvis, and keep the clamp for 30 minutes to maximize exposure; after clamping the renal artery\u002Fvein via the renal vein route, slowly inject 100 μL through the proximal end of the renal vein [12-50,12-51]. Coverage: AAV9 can transduce all nephron segments (proximal tubules, distal tubules, collecting ducts, podocytes), and can be segment-specific with segment-specific promoters (SGLT2→PT, etc.); extrarenal expression is minimal, and liver off-targeting is significantly lower than systemic administration; expression peaks in 2-4 weeks [12-50,12-51]. \u003C\u002Fp>","2026-09-18 11:24:28","2026-10-18 02:37:45",{"id":191,"service_pic":7,"service_title":192,"service_class":101,"class_name":102,"service_content":193,"service_tag":104,"created_at":194,"updated_at":195},33,"Pancreatic Duct Injection (Pancreatic Duct)","\u003Cp>Applicable species: mice; there are few data on rats\u002FNHP (NHP is mainly explored by ERCP) [12-52,12-53]. Dose range: 2×10^11-2×10^12 vg\u002Fmouse (AAV6) [12-52,12-53]. Injection volume: 100 μL\u002Fanimal [12-52,12-53]. Needle: Intubation through duodenal papilla\u002Fcommon bile duct (usually 30 G or finer, to be cored) [12-52,12-53]. Speed: 10 μL\u002Fmin×10 min=100 μL (Peristaltic P33 peristaltic pump) [12-52,12-53]. Anesthesia: isoflurane inhalation + oxygen, postoperative buprenorphine 0.075 mg\u002Fkg for analgesia [12-52,12-53]. Operational points: Make a midline incision in the abdomen to expose the duodenum and common bile duct. The proximal end of the common bile duct is clamped with bulldog clips to prevent viruses from entering the liver. The duodenal papilla is punctured and cannulated into the common bile duct\u002Fpancreatic duct. 100 μL AAV6 is slowly perfused with a peristaltic pump. After perfusion, the clips are released and the abdomen is closed layer by layer [12-52,12-53]. Coverage: AAV6 has the highest transduction of acinar cells (about 37% acinar expression), ductal cells are also transduced, and islet cell transduction is limited; the Ela promoter can be acinar-specific; the peak expression is 4-6 weeks [12-52,12-53]. Safety: Too fast perfusion (>50 μL\u002Fmin) or too high pressure can induce acute pancreatitis, and the peristaltic pump must be accurately controlled; AAV6 itself does not cause chronic pancreatitis, but the operation has the risk of mild inflammation [12-52,12-53]. \u003C\u002Fp>","2026-09-18 11:24:48","2026-10-19 00:23:11",{"id":197,"service_pic":7,"service_title":198,"service_class":101,"class_name":102,"service_content":199,"service_tag":104,"created_at":200,"updated_at":201},34,"Intra-Articular (IA) injection","\u003Cp>Applicable species: mice (knee\u002Fankle), rats (knee), rabbits, horses [12-54, 12-55, 12-63]. Dose range: mice 0.5×10^10-1×10^11 vg\u002Fjoint; rats 1×10^10-2.5×10^10 vg\u002Fjoint; horses refer to clinical OA gene therapy levels [12-54, 12-55, 12-63]. Injection volume: 5 μL\u002Fjoint for mice; 50 μL\u002Fjoint for rats [12-54, 12-55, 12-63]. Needle: mouse Hamilton+30 G needle; rat Hamilton+30 G or 27 G [12-54, 12-55, 12-63]. Speed: After slow injection, gentle flexion and extension of the joints can help distribution [12-54, 12-55, 12-63]. Anesthesia: Ketamine + xylazine IP or isoflurane [12-54, 12-55, 12-63]. Key points of operation: The needle is inserted at the medial edge of the patellar tendon of the mouse knee joint and the needle is inserted at the anterolateral aspect of the ankle joint; the disappearance of tactile resistance in rats indicates entry into the joint cavity, with a maximum volume of 35-50 μL [12-54, 12-55, 12-63]. Coverage: mainly transduces synovial lining cells (fibroblast-like synovial cells), with very limited transduction of chondrocytes (cartilage matrix barrier); expression can last for several months (slow synovial cell renewal); the equine OA model AAV2.5-IL-1Ra achieves sustained intra-articular expression [12-54, 12-55, 12-63]. Limitations: Treatment of cartilage lesions requires direct intrachondral injection\u002Fcapsid engineering optimization\u002Fparasynovial secretion, and simple IA cannot penetrate cartilage [12-54, 12-55, 12-63]. \u003C\u002Fp>","2026-09-18 11:25:05","2026-10-28 15:14:27",{"id":203,"service_pic":7,"service_title":204,"service_class":101,"class_name":102,"service_content":205,"service_tag":104,"created_at":206,"updated_at":207},35,"Intratumoral (ITu)","\u003Cp>Applicable species: mice (subcutaneous\u002Fin situ tumors such as 4T1, RENCA, B16, etc.), rats, and dogs with spontaneous tumors [12-64]. Dose range: mice 5×10^10-1×10^11 vg\u002Ftumor; rats 1×10^10-5×10^10 vg\u002Ftumor [12-64]. Injection volume: mice 50-100 μL\u002Ftumor (single point or 2-4 points); rats 100-200 μL[12-64]. Needle: 30 G needle percutaneous ultrasound guidance or Hamilton [12-64]. Anesthesia: Isoflurane inhalation. Operational points: Use ultrasound to confirm the tumor boundary and necrotic area (avoid the necrotic area), puncture the 30 G needle to the center of the tumor biopsy and inject slowly, which can cover different areas at multiple points [12-64]. Coverage: The transduction of tumor cells around the injection point decreases with distance, which can cause changes in the local immune microenvironment; off-target leakage to surrounding normal tissues or intravascular leakage to the liver\u002Fsystem [12-64]. \u003C\u002Fp>","2026-09-18 11:25:21","2026-10-19 14:01:53",{"id":209,"service_pic":7,"service_title":210,"service_class":101,"class_name":102,"service_content":211,"service_tag":104,"created_at":212,"updated_at":213},36,"Sciatic\u002FPeripheral Nerve Injection","\u003Cp>Applicable species: mice, rats, NHP (cynomolgus monkeys) [12-56,12-57]. Dose range: mouse 2.1×10^8 vg (scAAV1, 2 μL divided into 2 branches); rat 2×10^11 GC\u002Fnerve; NHP single nerve 2.5×10^12-2.6×10^13 vg\u002Fnerve (3×10^12-2.1×10^13 vg\u002Fkg based on body weight) [12-56,12-57]. Injection volume: 2 μL for mice; 20 μL for rats; 300 μL bolus for NHP each time[12-56,12-57]. Needle: mouse glass microelectrode\u002FHamilton; NHP ultrasound-guided nerve block blunt needle + nerve stimulator confirmation (≥0.3 mA indicates that it is safer to be farther away from the fiber) [12-56,12-57]. Speed: NHP 100 μL\u002Fmin, the distance between adjacent injection points is 3 cm (300 μL spreads about 3 cm) [12-56,12-57]. Operating points: NHP ultrasound locates nerves, and each animal is injected with 10-20 nerves; it can contain 0.1% Fast Green for visualization [12-56,12-57]. Coverage: AAV2\u002F9 can penetrate the perineurium and transduce up to 90% of myelinated Schwann cells, evenly distributed along the 15 cm nerve length; scAAV1 is most efficiently transported retrogradely to DRG and spinal motor neurons; there are off-targets in the liver (0.9 vg\u002Fdg) and spleen (3.6 vg\u002Fdg) [12-56,12-57]. Safety: Excessive intraneural injection pressure can cause capillary collapse\u002Fischemia\u002Faxonal damage. NHP recommends the perineural route as the first choice for clinical translation [12-56,12-57]. \u003C\u002Fp>","2026-09-18 11:25:44","2026-10-19 11:39:51",{"id":215,"service_pic":7,"service_title":216,"service_class":101,"class_name":102,"service_content":217,"service_tag":104,"created_at":218,"updated_at":219},37,"Cochlea\u002FInner Ear Injections (Cochlea\u002FInner Ear)","\u003Cp>Applicable species: mice (P0-P30 are more efficient), rats, guinea pigs, NHP[12-58]. Dose range: mouse 1×10^10-5×10^10 vg\u002Fear; NHP 1×10^11-7×10^11 vg\u002Fear[12-58]. Injection volume: mouse 0.6-2 μL\u002Fear; NHP 10-40 μL\u002Fear (perilymph volume is approximately 26.7 μL); guinea pig 2-10 μL (perilymph 8.66 μL) [12-58]. Needle: mouse borosilicate glass microelectrode (tip 20-40 μm) or 33 G needle; NHP 33 G trans-round window membrane [12-58]. Speed: mouse 0.1-0.5 μL\u002Fmin; NHP 20-100 nL\u002Fs[12-58]. Anesthesia: hypothermic anesthesia for neonatal rats, isoflurane for adults; NHP general anesthesia + postoperative cefazolin\u002Fanalgesia [12-58]. Key points of the operation: Make an incision behind the ear to expose the auditory bubble, grind open to expose the round window niche (RW niche), puncture the center of the RWM with a glass microelectrode, slowly inject after 5-10 minutes of stable perilymph outflow, and seal the RWM with muscle + tissue glue after injection; full cochlear transduction can be achieved in adult mice by combining posterior semicircular canal fenestration (RWM + CF) [12-58]. Coverage: RWM injection mainly transduces basal hair cells, combined with PSCC windowing, 94.6% apical transduction, 96.8% intermediate transduction, and basal transduction 94.2% of inner hair cells; high-dose almost 100% IHC\u002FOHC transduction; expression peaks in 2-4 weeks [12-58]. Safety: The injection volume does not exceed the perilymph volume (0.62 μL for mice, 26.7 μL for NHP, and 51 μL for humans), and the injection speed must be extremely slow to avoid hearing damage; overdose can overflow into the posterior cranial fossa through the cochlear aqueduct and cause brain transduction [12-58]. \u003C\u002Fp>","2026-09-18 11:26:07","2026-10-18 03:13:55",{"id":221,"service_pic":7,"service_title":222,"service_class":101,"class_name":102,"service_content":223,"service_tag":104,"created_at":224,"updated_at":225},38,"Intramyocardial injection","\u003Cp>Applicable species: mice, rats, NHP, dogs, pigs [12-59]. Dose range: 5×10^10-1×10^11 vg\u002Fheart for mice; 1×10^11-5×10^11 vg\u002Fheart for rats; approximately 10^11-10^12 vg level for pigs based on body weight (to be verified) [12-59]. Injection volume: a total of 30 μL divided into 3 points for mice (10 μL for each ventricular\u002Fdorsal\u002Flateral side of the left ventricular wall); 30-50 μL for rats; 100-200 μL for each point in pigs [12-59]. Needles: mouse Hamilton+31 G needle; canine 7-Fr endocardial 27 G injection needle catheter (Stiletto); porcine 30 G tip curved needle for thoracotomy and direct vision [12-59]. Anesthesia: isoflurane for mice (quick operation with small incision without mechanical ventilation); general anesthesia + mechanical ventilation for rats\u002Fdogs\u002Fpig[12-59]. Key points of operation: Thoracotomy through a small incision in the fourth intercostal space of mice, 10 μL at each of three points on the left ventricular wall after externalization of the heart; canine fluoroscopy + MRI fusion imaging (XFM) guides the catheter into the left ventricle through the aortic valve for multi-point injection; pigs are exposed through thoracotomy and injected at multiple points at the apex\u002Finfarct area\u002Fborder area [12-59]. Coverage: focal transduction of cardiomyocytes around the injection point, not covering the whole heart; peak expression in 3-4 weeks; dystrophin can be restored in DMD models [12-59]. Off-target: drug leakage from the pericardium\u002Fthoracic cavity, if injected into the left ventricular cavity, the drug solution will enter the circulation and cause liver off-target. \u003C\u002Fp>","2026-09-18 11:26:25","2026-10-21 23:57:43",{"id":227,"service_pic":7,"service_title":228,"service_class":101,"class_name":102,"service_content":229,"service_tag":104,"created_at":230,"updated_at":231},39,"Fat Pad Injection (Fat Pad)","\u003Cp>Applicable species: mice (eWAT, iBAT, iWAT); rats to be nucleated[12-60]. Dose range: mouse 1×10^11-2×10^11 vg\u002Fpad (AAV8), BAT about 1×10^9-2×10^10 vg\u002Fpad [12-60]. Injection volume: eWAT 30-50 μL\u002Fpad divided into 2 points; iBAT 20 μL\u002Fleaf (a total of 40 μL for both leaves); iWAT 25 μL\u002Fside[12-60]. Needle: Hamilton+30 G needle, BAT with 32 G fine injection, or BD insulin needle 31 G 8 mm [12-60]. Anesthesia: Isoflurane 2.5% for induction and 1% for maintenance [12-60]. Operational points: eWAT is injected at multiple points after the midline abdominal incision is exposed; iBAT is injected into two lobes through the interscapular longitudinal incision; inject slowly to confirm that there is no leakage; each point of eWAT is ≤10 μL to prevent leakage into the abdominal cavity [12-60]. Coverage: mature adipocytes + stromal vascular components, AAV8 is the most efficient in transducing adipose tissue, and the ADIPOQ promoter is specific for mature adipocytes; expression lasts for >4 weeks [12-60]. Off-target: intra-abdominal leakage (eWAT), high-dose hepatic spread. \u003C\u002Fp>","2026-09-18 11:26:48","2026-10-19 14:10:29",{"id":233,"service_pic":7,"service_title":234,"service_class":101,"class_name":102,"service_content":235,"service_tag":104,"created_at":236,"updated_at":236},40,"Intradermal\u002FWound injection (Intradermal\u002FWound)","\u003Cp>Applicable species: mice, rats, pigs (skin structure is highly similar to humans) [12-61]. Dose range: The working concentration of mice is 1×10^12 vg\u002FmL, each point is about 2×10^10 vg, and the total dose is 1×10^10-1×10^11 vg\u002Fwound [12-61]. Injection volume: 10-20 μL per point, 4-6 points per wound, 80-120 μL in total; 50-100 μL per point in rats [12-61]. Needle: Insulin syringe 29-31 G or Hamilton [12-61]. Anesthesia: Ketamine 80 mg\u002Fkg + xylazine 10 mg\u002Fkg IP or isoflurane [12-61]. Key points of operation: The tip of the intradermal injection needle is inserted into the skin at an angle of 10-15°, and the wheal can be seen when the needle is injected just after passing through the epidermis; the wound should be injected intradermally at 4-6 points around the wound edge or dripped directly into the wound bed and then covered with Vaseline gauze\u002Fhydrogel stent [12-61]. Coverage: Epidermal AAV2\u002F6 can transduce basal keratinocytes (KRT14 promoter), dermal fibroblasts, and wound repair cells; expression peaks in 2-4 weeks [12-61]. Note: The intact stratum corneum of the skin is the main barrier for transdermal penetration, and intradermal injection is limited to a few millimeters around the injection point. \u003C\u002Fp>","2026-09-18 11:27:12",{"id":238,"service_pic":7,"service_title":239,"service_class":92,"class_name":93,"service_content":240,"service_tag":95,"created_at":241,"updated_at":242},41,"Cross-species dose conversion methodology","\u003Cp>AAV dose conversion is not a single mathematical formula, but a structured scientific argument that combines administration route, target organ size, species transduction efficiency, immunobiology and product quality. Simply equating mouse vg\u002Fkg directly to human vg\u002Fkg is a common mistake because target organ\u002Fcell numbers are not proportional to body weight, and there are species differences in AAV receptor biology and glycan presentation [12-41]. \u003C\u002Fp>\n\u003Cp>Conversion path one: body weight conversion (vg\u002Fkg), suitable for systemic\u002Fintravenous administration. In clinical IV administration, administration is usually based on body weight (vg\u002Fkg = total vg\u002Fkg body weight). The AAV field linearly converts based on body weight and does not use body surface area (BSA) - this is a key difference from traditional small molecule\u002Fantibody drugs. Basis: AAV liver\u002Fheart\u002Fmyocardial transduction is reported in vg\u002Fkg in large animals, and the SMA clinical Zolgensma dose of 1.1×10^14 vg\u002Fkg is directly extrapolated from NHP vg\u002Fkg data [12-11,12-41]. \u003C\u002Fp>\n\u003Cp>Conversion path two: organ weight\u002Fvolume conversion, suitable for local\u002Fintra-organ injection. When injecting into an organ with clear structural boundaries, it is converted based on organ weight or volume. Clinical local injection is usually based on a fixed dose (vg\u002Fsubject) or a fixed concentration (vg\u002FmL\u002Fsubject). Examples of eye vitreous volume conversion: mouse 0.0044 mL, dog 3 mL, NHP 2 mL, human 4 mL [12-44]. \u003C\u002Fp>\n\u003Cp>Conversion path three: Allometric Scaling, for auxiliary reference only. The classic formula Y=aW^b is based on body surface area normalization and is only a supportive reference for AAV and cannot replace species-specific biodistribution, efficacy, toxicology and potency data [12-42]. \u003C\u002Fp>\n\u003Cp>Conversion path four: Activity Factor corrects cross-species transduction efficiency. The composite metric represents expected interspecies differences in transduction and transgene expression between animal models and humans. Example: scAAV2\u002F8-LP1-hFIXco mice → NHP peak FIX levels decreased 20-fold, NHP → humans decreased 3-fold; Valoctocogene roxaparvovec (AAV5) NHP to human dose-adjusted peak ratio was 0.38 [12-43]. \u003C\u002Fp>\n\u003Cp>The conversion difference between systemic IV vs local\u002Forgan injection: System IV is linearly converted according to vg\u002Fkg; local injection is converted according to organ volume\u002Ffixed dose, and linear scaling of body weight is not applicable. Extrapolation between species needs to be corrected: capsid receptor differences (AAV-PHP.B\u002FLy6a is only effective for C57BL\u002F6J), pre-existing neutralizing antibodies (NHP\u002Fhuman prevalence is high), opposite liver zone distribution (mouse periportal vs NHP pericentral), high-dose toxicity threshold NHP\u002Fpig is more sensitive than mice (mice tolerate >1×10^14 vg\u002Fkg, NHP is about 2×10^14 GC\u002Fkg means severe toxicity) [12-7,12-11,12-12,12-15]. Mice → NHP experience conversion: mice commonly use 1×10^12 vg\u002F20 g = 5×10^13 vg\u002Fkg, NHP equivalent starts at 2×10^12 vg\u002Fkg and then ramps up according to effect\u002Ftoxicity [12-11]. \u003C\u002Fp>\n\u003Cp>Conversion precautions: The titer determination method, empty\u002Fsolid shell ratio, potency determination and vector preparation comparability must be verified before using the literature dose; scAAV and ssAAV cannot use universal potency multipliers, and data need to be directly compared; large differences in dosage between newborn animals and adult animals need to be reported separately; FDA\u002FEMA do not provide fixed conversion formulas and require scientific demonstration based on product specificity. High-dose system AAV must have NHP or large animal biodistribution and toxicology data [12-41,12-43]. \u003C\u002Fp>","2026-09-18 11:28:14","2026-10-22 08:59:56",{"id":244,"service_pic":7,"service_title":245,"service_class":92,"class_name":93,"service_content":246,"service_tag":95,"created_at":247,"updated_at":248},42,"High dose safety tips","\u003Cp>Systemic IV high-dose hepatotoxicity\u002Fthrombotic microangiopathy (TMA): NHP and piglet IV 2×10^14 GC\u002Fkg AAVhu68. All 3 NHPs had elevated transaminases, and 1 had acute liver failure + shock and was euthanized on the 4th day. Severe toxicity was also seen with piglet; ≥1×10^14 Hepatic sinusoidal platelet microthrombi and endothelial injury (SOS) at GC\u002Fkg have nothing to do with transgene; thrombocytopenia, coagulation abnormalities, and complement activation occurred 1-2 weeks after administration, which are related to pre-existing anti-capsid antibodies [12-11,12-25]. \u003C\u002Fp>\n\u003Cp>CSF administration DRG lesions: DRG mononuclear cell infiltration, neuronal degeneration, and axonopathy after NHP administration through blood or CSF; 33 NHPs were collected, 83% of the CSF pathways were involved, and they were usually clinically silent. The dose threshold is approximately ≥1×10^13 vg\u002FIT\u002FICM, and \u003C5×10^12 vg\u002Funit is usually asymptomatic; elevated plasma\u002FCSF NfL indicates DRG damage; the mitigation strategy is to insert a miRNA target sequence into the expression cassette to target the DRG transgene [12-26, 12-27, 12-36]. \u003C\u002Fp>\n\u003Cp>Clinical death signal: DMD high-dose IV (approximately 2×10^14 vg\u002Fkg level) rAAV9 causes liver failure, thrombotic microangiopathy, and death (Lek 2023 NEJM) [12-28]. \u003C\u002Fp>\n\u003Cp>Restrictions on immunogenicity and repeated administration: AAV induces anti-capsid neutralizing antibodies (NAb), which strongly limits repeated administration. Systemic administration with multiple injection intervals of ≥2 weeks is still restricted by NAb; strong promoter + neoantigen can cause cellular immune clearance in immune-competent animals, which is more obvious in NHP\u002Fhuman, and immune regulation (such as rapamycin SVP) can be evaluated; NHP IM AAV8 shows a loss of immune-mediated expression but is not irreversible [12-12,12-24]. \u003C\u002Fp>\n\u003Cp>Mouse strain barrier and others: AAV-PHP.B strong BBB penetration is limited to C57BL\u002F6J (Ly6a dependent), BALB\u002Fc and other strains do not have this phenotype [12-7]; high-dose ubiquitous promoter (CBA\u002FCMV) can induce neuronal stress and microgliosis in the target area; pig AAV8-IL-10 IT has caused fatal meningitis, suggesting that the immunogenicity of the transgenic product itself needs to be evaluated separately [12-34]. \u003C\u002Fp>","2026-09-18 11:28:32","2026-10-29 02:08:46",{"id":250,"service_pic":7,"service_title":251,"service_class":92,"class_name":93,"service_content":252,"service_tag":95,"created_at":253,"updated_at":254},43,"Reference Doses and Detection Time Points for AAV Studies","\u003Cp>It is recommended to set early (1-2 weeks, confirm the onset of effect), primary endpoint (3-6 weeks, compare expression and distribution) and sustained endpoint (8-12 weeks or longer). Detection should include at least two layers of vector DNA, transgenic mRNA, and protein\u002Ffunctional readout; for tissue distribution issues, non-target tissue and empty vector\u002Fnegative controls should be added. For actively proliferating tissues, long-term expression decline may result from vector genome dilution rather than batch quality issues. [12-2,12-3]\u003C\u002Fp>","2026-09-18 11:28:46","2026-10-19 14:11:41",{"id":256,"service_pic":7,"service_title":79,"service_class":92,"class_name":93,"service_content":257,"service_tag":95,"created_at":258,"updated_at":258},44,"\u003Cp>[12-1] Challis RC, et al. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nat Protoc. 2019;14:379-414. DOI:10.1038\u002Fs41596-018-0097-3. [H]\u003C\u002Fp>\n\u003Cp>[12-2] Hudry E, Vandenberghe LH. Therapeutic AAV gene transfer to the nervous system. Neuron. 2019;102:263-271. DOI:10.1016\u002Fj.neuron.2019.03.020. [H]\u003C\u002Fp>\n\u003Cp>[12-3] Kavita U, et al. 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Robust spinal motor neuron transduction following intrathecal delivery of AAV9 in pigs. Gene Ther. 2012;19(8):852-859. DOI:10.1038\u002Fgt.2011.187. [H]\u003C\u002Fp>\n\u003Cp>[12-36] Hordeaux J, Hinderer C, Goode T, et al. Toxicology study of intra-cisterna magna AAV9 expressing iduronate-2-sulfatase in rhesus macaques. Mol Ther Methods Clin Dev. 2018;10:68-78. DOI:10.1016\u002Fj.omtm.2018.06.004. PMID:30073178. [N]\u003C\u002Fp>\n\u003Cp>[12-37] Samaranch L, Salegio EA, San Sebastian W, et al. AAV9 transduction in the central nervous system of nonhuman primates. Hum Gene Ther. 2012;23(4):382-389. DOI:10.1089\u002Fhum.2011.141. [H]\u003C\u002Fp>\n\u003Cp>[12-38] Golebiowski D, et al. Direct intracranial injection of AAVrh8 encoding monkey beta-N-acetylhexosaminidase causes neurotoxicity in the primate brain. Hum Gene Ther. 2017;28(7):510-522. DOI:10.1089\u002Fhum.2016.109. [N]\u003C\u002Fp>\n\u003Cp>[12-39] Haurigot V, Marco S, Ribera A, et al. Whole body correction of mucopolysaccharidosis IIIA by intracerebrospinal fluid gene therapy. J Clin Invest. 2013;123(8):3254-3271. DOI:10.1172\u002FJCI66778. [H]\u003C\u002Fp>\n\u003Cp>[12-40] AAV2-mediated gene delivery to monkey putamen: infusion device and delivery parameters. PMC3816113. [H]\u003C\u002Fp>\n\u003Cp>[12-41] Tang F, et al. Rational clinical dose selection of AAV-mediated gene therapy based on allometric principles. Clin Pharmacol Ther. 2021;110(3):803-807. DOI:10.1002\u002Fcpt.2269. [N]\u003C\u002Fp>\n\u003Cp>[12-42] Nair AB, Jacob S. A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm. 2016;7(2):27-31. DOI:10.4103\u002F0976-0105.177704. PMID:27057118. [H]\u003C\u002Fp>\n\u003Cp>[12-43] The perspective of DMPK on rAAV-based gene therapy. AAPS J. 2022. PMC8817103. [N]\u003C\u002Fp>\n\u003Cp>[12-44] Clinical pharmacology perspective on AAV retina gene therapy. Clin Pharmacol Ther. 2024. DOI:10.1002\u002Fcpt.3230. 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AAV vector distribution in the mouse respiratory tract following four different methods of administration. BMC Biotechnol. 2017;17:43. DOI:10.1186\u002Fs12896-017-0365-2. [H]\u003C\u002Fp>\n\u003Cp>[12-50] Rocca CJ, et al. rAAV9 combined with renal vein injection is optimal for kidney-targeted gene delivery. Gene Ther. 2014;21:618-628. DOI:10.1038\u002Fgt.2014.35. [F]\u003C\u002Fp>\n\u003Cp>[12-51] Asico LD, et al. Nephron segment-specific gene expression using AAV vectors. Biochem Biophys Res Commun. 2018;497:19-24. DOI:10.1016\u002Fj.bbrc.2018.01.178. [H]\u003C\u002Fp>\n\u003Cp>[12-52] Quirin KA, et al. AAV retrograde pancreatic ductal gene delivery. Mol Ther Methods Clin Dev. 2018;8:8-20. DOI:10.1016\u002Fj.omtm.2017.10.004. [H]\u003C\u002Fp>\n\u003Cp>[12-53] Xiao X, Guo P, Prasadan K. Pancreatic cell tracing, lineage tagging and targeted genetic manipulations using pancreatic ductal infusion of AAV vectors. Nat Protoc. 2014;9:2719-2724. DOI:10.1038\u002Fnprot.2014.183. [F]\u003C\u002Fp>\n\u003Cp>[12-54] Xing Y, et al. Intra-articular AAV-mediated PRG4 gene therapy for preventing posttraumatic osteoarthritis. Mol Ther. 2022;30:1864-1876. DOI:10.1016\u002Fj.ymthe.2022.02.015. [N]\u003C\u002Fp>\n\u003Cp>[12-55] Frisch J, et al. Persistence, localization, and external control of transgene expression after single injection of AAV into injured joints. Hum Gene Ther. 2013;24:423-433. DOI:10.1089\u002Fhum.2012.118. [H]\u003C\u002Fp>\n\u003Cp>[12-56] Carré A, et al. Perineural delivery of AAV2\u002F9 in non-human primates is a safe and efficient route for gene therapy in Charcot-Marie-Tooth diseases. Mol Ther Methods Clin Dev. 2025. PMC12433476. [N]\u003C\u002Fp>\n\u003Cp>[12-57] Hutson MR, et al. Efficient retrograde neuronal transduction utilizing self-complementary AAV1. Mol Ther. 2012;20:1298-1307. [H]\u003C\u002Fp>\n\u003Cp>[12-58] Lv J, et al. AAV1-hOTOF gene therapy for autosomal recessive deafness 9. Lancet. 2024;403:2317-2325. DOI:10.1016\u002FS0140-6736(23)02874-X. PMID:38280389. [N]\u003C\u002Fp>\n\u003Cp>[12-59] Gao F, et al. A simple and efficient method for in vivo cardiac-specific gene manipulation by intramyocardial injection in mice. J Vis Exp. 2015;(102):e53022. PMC5933510. [F]\u003C\u002Fp>\n\u003Cp>[12-60] Jimenez V, et al. In vivo AAV-mediated genetic engineering of white and brown adipose tissue in adult mice. Diabetes. 2013;62:4012-4022. DOI:10.2337\u002Fdb13-0496. [F]\u003C\u002Fp>\n\u003Cp>[12-61] Keswani SG, et al. AAV vector tropism in murine wound healing. Wound Repair Regen. 2012;20:592-600. DOI:10.1111\u002Fj.1524-475X.2012.00817.x. [F]\u003C\u002Fp>\n\u003Cp>[12-62] Acland GM, Aguirre GD, Ray J, et al. Gene therapy restores vision in a canine model of childhood blindness. Nat Genet. 2001;28:92-95. DOI:10.1038\u002F88357. PMID:11385066. [H]\u003C\u002Fp>\n\u003Cp>[12-63] A phase 1 clinical trial shows safe, sustained AAV-mediated expression of IL-1Ra in human OA knee. Sci Transl Med. 2025. DOI:10.1126\u002Fscitranslmed.adu9804. [N]\u003C\u002Fp>\n\u003Cp>[12-64] Teo Q, et al. Inducible caspase 9-mediated suicide gene therapy using AAV6 in murine breast cancer. Mol Ther Oncolytics. 2023;30:187-198. [N]\u003C\u002Fp>","2026-09-18 11:29:20",{"id":260,"service_pic":7,"service_title":261,"service_class":6,"class_name":9,"service_content":262,"service_tag":11,"created_at":263,"updated_at":264},45,"General Principles for Tissue Application","\u003Cp>Tissue selection adopts the combined model of \"capsid entry × administration reachability × transcriptional regulation × dose × detection endpoint\". The serotypes and doses in the table below are common starting points for preliminary experiments in mice; changes in disease model, age, and manufacturing processes may alter results. \u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028044\">\u003C\u002Fa>Tissue serotype and promoter quick check\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>\u003Cstrong>Organization\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>\u003Cstrong>Common natural\u002Fengineered candidates\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>\u003Cstrong>Commonly used transcriptional controls\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>\u003Cstrong>Common Paths\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Retina\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV2, AAV5, AAV8 and eye engineering capsids\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>RPE65\u002FVMD2, GRK1, SNCG\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Subretinal, vitreous cavity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Liver\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV8, AAV5, AAV3B\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>TBG, hAAT, LP1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Veins\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Heart\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV9, AAV6\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>cTnT\u002FTNNT2, MYL2, MHCK7\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Multiple points in veins and myocardium\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Skeletal muscle\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV9, AAVrh74, MyoAAV\u002FAAVMYO\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>MHCK7, desmin, CK8\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Intravenous, intramuscular\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Bone\u002FCartilage\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV9, AAV-DJ or capsid screening\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>COL1A1, BGLAP, COL2A1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Local, intravenous\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Lungs\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV5, AAV6, AAV9 and lung-targeting capsids\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>SFTPC, SCGB1A1, FOXJ1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Intranasal, intratracheal, intravenous\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>kidney\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV9, AAV2\u002F8 (path dependency)\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>NPHS1\u002F2, CDH16, AQP2\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Renal vein, retrograde ureter\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Pancreas\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV6, AAV8, AAV9\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>INS1\u002F2, GCG, PDX1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Pancreatic duct, vein\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Fat\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV8, AAV9 and capsid screening\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>ADIPOQ, UCP1, FABP4\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Fat pad, vein\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"72\">\n\u003Cp>Skin\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>AAV2, AAV6, AAV8\u002F9\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"192\">\n\u003Cp>KRT14, KRT5, IVL, COL1A1\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"130\">\n\u003Cp>Intradermal, local wound area\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>","2026-09-18 11:29:45","2026-10-18 18:02:43",{"id":266,"service_pic":7,"service_title":267,"service_class":268,"class_name":269,"service_content":270,"service_tag":271,"created_at":272,"updated_at":273},47,"Retina",46,"Tissue-Specific Targeted Delivery Strategies","\u003Ch3>1.1 Retinal tissue structure\u003C\u002Fh3>\n\u003Cp>The retina is obviously stratified, and the access paths of the outer photoreceptor cells, RPE and inner ganglion cells are different. \u003C\u002Fp>\n\u003Ch3>1.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV2 is often used for inner layer\u002Fvitreous related design; AAV5, AAV8 and engineered capsids are often used for outer layer or subretinal delivery. \u003C\u002Fp>\n\u003Ch3>1.3 Retina-specific promoter\u003C\u002Fh3>\n\u003Cp>VMD2\u002FRPE65 is selected for RPE, GRK1 is selected for photoreceptor cells, and SNCG can be assessed for ganglion cells. \u003C\u002Fp>\n\u003Ch3>1.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Subretinal (SR): the gold standard pathway targeting the RPE and photoreceptors. Mouse 1×10^8-3×10^9 vg\u002Feye, 1-2 μL, 33 G Hamilton blunt needle, slowly inject 1-2 μL\u002Fmin and retain the needle for 30 s; NHP 7×10^11-7×10^12 vg\u002Feye (AAV8), 100-200 μL, 39 G injection cannula\u002F36-42 G retinal needle, AAV and Healon Mix 1:1 to form bleb; dog 1.0-1.4×10^11 vg\u002Feye, 200 μL; pig 1×10^10-1.25×10^11 vg\u002Feye [1-4, 1-5, 1-8]. \u003C\u002Fp>\n\u003Cp>Intravitreal cavity (IVT): Targeting RGCs and inner retina. Mouse 1×10^9-1×10^10 vg\u002Feye, 1-2 μL, 33 G blunt needle; rat 1-5×10^9 vg\u002Feye, 5 μL; NHP 7×10^11 vg\u002Feye, 100 μL, 30 G via pars plana. The internal limiting membrane (ILM) is the main barrier for IVT transduction of the outer retina, and engineering capsids such as AAV2.7m8 or ILM removal can improve penetration [1-7,1-8]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: SR only transduces RPE+ photoreceptors in the bleb area (the mouse bleb covers about 20-40% of the retina), and the off-target is extremely low; IVT is mainly RGC\u002Finner layer, and large animals have limited penetration of the outer retina. The peak expression is 3-6 weeks on average, and SR can last for >1.5 years. The transduction efficiency and immunogenicity of SR in NHP are better than those in IVT [1-5,1-6]. \u003C\u002Fp>\n\u003Ch3>1.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Acland et al. confirmed that AAV subretinal injection restored vision in the RPE65 mutant dog model, laying the foundation for RPE65 gene therapy [1-4]; Yiu et al. established the AAV2\u002F8 photoreceptor transduction dose threshold in monkeys (7×10^12 vg\u002Feye for extensive transduction, 7×10^11 vg\u002Feye (no obvious expression) [1-5]; The RD-CURE Alliance compared SR and IVT in NHP, and the retinal gene transfer and biodistribution of SR were significantly better than that of IVT [1-6]; Dalkara et al. demonstrated that ILM is a physical barrier to intravitreal transduction [1-7]. The design should report cell layer, transduction area, functional endpoints, and intraocular inflammation simultaneously. \u003C\u002Fp>\n\u003Ch3>1.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[1-1] Russell S, et al. Voretigene neparvovec phase 3 trial. Lancet. 2017;390:849-860. DOI:10.1016\u002FS0140-6736(17)31868-8. PMID:28712537. [H]\u003C\u002Fp>\n\u003Cp>[1-2] Fischer MD, et al. Subretinal gene therapy targeting cone photoreceptors. JAMA Ophthalmol. 2020;138:643-651. [H]\u003C\u002Fp>\n\u003Cp>[1-3] He X, et al. AAV for gene therapy in ocular diseases. Research. 2023;6:0291. [N]\u003C\u002Fp>\n\u003Cp>[1-4] Acland GM, Aguirre GD, Ray J, et al. Gene therapy restores vision in a canine model of childhood blindness. Nat Genet. 2001;28:92-95. DOI:10.1038\u002F88357. PMID:11385066. [H]\u003C\u002Fp>\n\u003Cp>[1-5] Yiu G, et al. Dosage thresholds for AAV2 and AAV8 photoreceptor gene therapy in monkey. Sci Transl Med. 2011;3:88ra54. DOI:10.1126\u002Fscitranslmed.3002103. PMID:21849668. [H]\u003C\u002Fp>\n\u003Cp>[1-6] RD-CURE Consortium. Superior retinal gene transfer and biodistribution profile of subretinal versus intravitreal delivery of AAV8 in nonhuman primates. Invest Ophthalmol Vis Sci. 2017;58:5792-5801. DOI:10.1167\u002Fiovs.17-22473. PMID:29186476. [H]\u003C\u002Fp>\n\u003Cp>[1-7] Dalkara D, et al. Inner limiting membrane barriers to AAV-mediated retinal transduction from the vitreous. Mol Ther. 2009;17:2096-2102. DOI:10.1038\u002Fmt.2009.181. PMID:19773742. [H]\u003C\u002Fp>\n\u003Cp>[1-8] Boye SE, et al. Suprachoroidal and subretinal injections of AAV using transscleral microneedles for retinal gene delivery in nonhuman primates. Hum Gene Ther. 2019;30:1123-1134. DOI:10.1089\u002Fhum.2019.052. [N]\u003C\u002Fp>","AAV, Tissue-Specific Targeted Delivery Strategies","2026-09-18 11:30:47","2026-10-16 21:47:56",{"id":275,"service_pic":7,"service_title":276,"service_class":268,"class_name":269,"service_content":277,"service_tag":271,"created_at":278,"updated_at":279},48,"Liver","\u003Ch3>2.1 Liver tissue structure\u003C\u002Fh3>\n\u003Cp>Sinusoidal endothelium and hyperperfusion make the liver susceptible to intravenous administration of AAV, but hepatocytes, stellate cells, and endothelial cells cannot be confused with the same target. \u003C\u002Fp>\n\u003Ch3>2.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV8 is often used as a starting point in mice; AAV5 has clinical experience; AAV3B is often evaluated in human hepatocyte models. \u003C\u002Fp>\n\u003Ch3>2.3 Liver-specific promoter\u003C\u002Fh3>\n\u003Cp>Hepatocytes are commonly used with TBG, hAAT or LP1; non-hepatocyte targeting must include cell type verification. \u003C\u002Fp>\n\u003Ch3>2.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Systemic vein (IV): first choice for liver targeting. 1×10^11-1×10^12 vg\u002Fmouse (approximately 5×10^12-5×10^13 vg\u002Fkg), 100-300 μL, 27-30 G needle, slow infusion for 30-60 s; AAV8\u002F9 reaches 90%+ hepatocyte transduction with dose, and detected in 2-6 weeks [2-1,2-5]. \u003C\u002Fp>\n\u003Cp>Portal vein (PV): direct liver targeting. 1×10^10-1×10^12 vg\u002Fmouse, ≤100 μL, open abdominal puncture with 33 G needle, and inject slowly for 1-2 min. AAV8 can transduce 90-95% of hepatocytes through PV; the minimally invasive alternative is to inject 50 μL through the spleen parenchyma. Whole body off-target is significantly lower than IV, but laparotomy is costly and unrepeatable [2-4,2-6]. \u003C\u002Fp>\n\u003Cp>Intraperitoneal (IP): Less bioavailable than IV but with enhanced diaphragm\u002Fadipose\u002Fpancreatic transduction. 1×10^11-1×10^12 vg\u002Fmouse, 100-200 μL injected into the left lower abdomen; liver transduction is lower than IV. Note that there are species differences in the distribution of liver lobule zones (mouse perihilar vs NHP pericentral) [2-4,2-5]. \u003C\u002Fp>\n\u003Ch3>2.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Barcode amplicon sequencing systematically compares the administration route-dependent transduction profiles of commonly used AAV variants in mice, providing a quantitative basis for IV\u002FPV\u002FIP route selection [2-5]; retrograde zone distribution studies reveal that the transduction zones in mice and NHP liver lobes are opposite, and care must be taken when extrapolating hepatocyte expression across species [2-4]; The JoVE protocol details the operation of injecting AAV8-shRNA through the spleen to achieve efficient gene knockdown in the liver [2-6]. The species and previous neutralizing antibodies have obvious effects, and the expression of blood factors does not equal the complete recovery of liver cell function. \u003C\u002Fp>\n\u003Ch3>2.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[2-1] Wang D, et al. AAV vector as a platform for gene therapy delivery. Nat Rev Drug Discov. 2019. [H]\u003C\u002Fp>\n\u003Cp>[2-2] Pipe SW, et al. Etranacogene dezaparvovec for hemophilia B. N Engl J Med. 2023;388:706-718. DOI:10.1056\u002FNEJMoa2211644. PMID:36812434. [N\u002FH]\u003C\u002Fp>\n\u003Cp>[2-3] Ben-Moshe S, Itzkovitz S. Spatial heterogeneity in the mammalian liver. Nat Rev Gastroenterol Hepatol. 2019;16:395-410. [H]\u003C\u002Fp>\n\u003Cp>[2-4] Inverse zonation of hepatocyte transduction with AAV vectors between mice and non-human primates. Mol Ther. 2014. PMC3269907. [H]\u003C\u002Fp>\n\u003Cp>[2-5] Mapping administration route-dependent transduction profiles of commonly used AAV variants in mice by barcode amplicon sequencing. 2025. PMID:40337478. [N]\u003C\u002Fp>\n\u003Cp>[2-6] Efficient gene knockdown in liver via intrasplenic injection of shRNA delivered by AAV8. JoVE. 2024;(163):e67376. DOI:10.3791\u002F67376. [F]\u003C\u002Fp>","2026-09-18 11:31:07","2026-10-25 16:17:26",{"id":281,"service_pic":7,"service_title":282,"service_class":268,"class_name":269,"service_content":283,"service_tag":271,"created_at":284,"updated_at":285},49,"Heart","\u003Ch3>3.1 Heart tissue structure\u003C\u002Fh3>\n\u003Cp>Cardiomyocytes, fibroblasts, endothelial cells, and conduction systems have different expression needs. \u003C\u002Fp>\n\u003Ch3>3.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>Systemic drug delivery is often evaluated based on AAV9, and local myocardial injection can be compared with AAV6; cardiomyocyte specificity cannot be judged based on whole-heart fluorescence alone. \u003C\u002Fp>\n\u003Ch3>3.3 Cardiac-specific promoter\u003C\u002Fh3>\n\u003Cp>cTnT\u002FTNNT2, MYL2 or MHCK7 are commonly used in cardiac muscle; fibroblast projects should use independent regulatory elements and cell markers. \u003C\u002Fp>\n\u003Ch3>3.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Systemic vein (IV): AAV9 has natural tropism for myocardium. 1×10^11-1×10^12 vg\u002Fmouse, 100-300 μL; Inagaki confirmed that the global cardiac gene transfer of AAV9 mice after systemic administration is better than that of AAV8[3-4]. \u003C\u002Fp>\n\u003Cp>Local injection into the myocardium: focal myocardial transduction. Mouse 5×10^10-1×10^11 vg\u002Fheart, a total of 30 μL divided into 3 points (10 μL each ventricular\u002Fdorsal\u002Fside of the left ventricular wall), 31 G Hamilton, the fourth intercostal small incision was made through thoracotomy and the heart was externally injected; rat 1×10^11-5×10^11 vg\u002Fheart, 30-50 μL multiple points; dog 7-Fr endocardial 27 The G injection needle catheter enters the left ventricle through the aortic valve, and is guided by fluoroscopy + MRI fusion; pig thoracotomy is used for multi-point injection [3-5]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: IV myocardial transduction is extensive but accompanied by liver off-target; local myocardial transduction is focal around the injection point, not covering the whole heart, and is suitable for locating the infarct boundary zone. The expression peaks in 3-4 weeks, and dystrophin can be restored in the DMD model [3-5,3-6]. It is recommended to simultaneously evaluate cardiac function, tissue expression, inflammation, and liver off-targets. \u003C\u002Fp>\n\u003Ch3>3.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Gao et al. established a simple and efficient method for cardiac-specific gene manipulation by intramyocardial injection in mice [3-5]; MRI roadmap-guided transendocardial rAAV delivery restored dystrophin expression in the canine DMD model [3-6]; Inagaki et al. systematically compared the cardiac transduction efficiency of systemic administration of AAV9 and AAV8 [3-4]. \u003C\u002Fp>\n\u003Ch3>3.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[3-1] Litviňuková M, et al. Cells of the adult human heart. Nature. 2020;588:466-472. [H]\u003C\u002Fp>\n\u003Cp>[3-2] Wang J, et al. AAV6 versus AAV9 after intramyocardial administration. Mol Ther Methods Clin Dev. 2025;33:101532. [N]\u003C\u002Fp>\n\u003Cp>[3-3] Ravindran D, et al. High-throughput evaluation of cardiac-specific promoters for AAV gene therapy. Gene Ther. 2025. [N]\u003C\u002Fp>\n\u003Cp>[3-4] Inagaki K, et al. Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8. Mol Ther. 2006;14(1):45-53. DOI:10.1016\u002Fj.ymthe.2006.03.011. [H]\u003C\u002Fp>\n\u003Cp>[3-5] Gao F, et al. A simple and efficient method for in vivo cardiac-specific gene manipulation by intramyocardial injection in mice. J Vis Exp. 2015;(102):e53022. PMC5933510. [F]\u003C\u002Fp>\n\u003Cp>[3-6] MRI roadmap-guided transendocardial delivery of exon-skipping rAAV restores dystrophin in canine DMD model. PMC3424392. [H]\u003C\u002Fp>","2026-09-18 11:31:36","2026-10-19 13:43:36",{"id":287,"service_pic":7,"service_title":288,"service_class":268,"class_name":269,"service_content":289,"service_tag":271,"created_at":290,"updated_at":291},50,"Skeletal Muscle","\u003Ch3>4.1 Muscle tissue structure\u003C\u002Fh3>\n\u003Cp>The transduction of muscle fibers, satellite cells, smooth muscles and vascular pericytes varies greatly, and the local intramuscular results cannot directly represent the whole body muscle group. \u003C\u002Fp>\n\u003Ch3>4.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV9 and AAVrh74 can be used as natural candidates; MyoAAV and AAVMYO represent engineering directions for cross-model screening. \u003C\u002Fp>\n\u003Ch3>4.3 Muscle-specific promoter\u003C\u002Fh3>\n\u003Cp>MHCK7, desmin and CK8 are used for different strength and size balances; myocardial and liver expression needs to be monitored. \u003C\u002Fp>\n\u003Ch3>4.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Intramuscular (IM): Injection into muscle. Mouse single point: 1×10^9-1×10^11 vg\u002Fmuscle, 25-50 μL, 29-30 G Hamilton, slow bolus injection for 30-60 s; rat single point: about 1×10^11 vg\u002Fmuscle, 50 μL; NHP rhesus IM AAV8 3×10^12 GC\u002Fkg divided into bilateral vastus lateralis muscles, 0.5-1 mL\u002Fpoint, 100 points can reach 135 mL; canine GRMD system IV 0.5-6.24×10^14 vg\u002Fkg[4-4,4-5]. \u003C\u002Fp>\n\u003Cp>System IV: Covered by skeletal muscles throughout the body. Mouse 1×10^11-1×10^12 vg\u002Fmouse; AAV9\u002FMyoAAV type extensive skeletal muscle transduction. Multi-muscle strategy: Multiple points (TA, gastrocnemius, quadriceps) dispersed injection covers more muscle fibers, and the upper limit of the total dose is limited by liver toxicity; regional limb perfusion (VLP\u002FIAPD) is used to replace multi-point IM in large animals [4-6]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: IM only injects muscle transduction, not distal muscle, and the secreted transgene can enter the bloodstream; NHP IM AAV8 shows unexpected significant liver targeting (the number of liver GCs is equivalent to that of local muscle); expression lasts for 2-6 weeks, and terminal differentiation of muscle fibers lasts for ≥6-12 months. Advantages of engineered capsids must be verified in target strains and production batches [4-4,4-6]. \u003C\u002Fp>\n\u003Ch3>4.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Studies have shown that IM injection of AAV8 in mice and macaques is accompanied by significant liver targeting and transgene expression [4-4]; Kornegay et al. evaluated systemic AAV-microdystrophin gene therapy in GRMD canine models [4-5]; bridging studies optimized NHP regimens from IM to limb perfusion delivery [4-6]. Loss of IM AAV expression in NHP is immune-mediated but not irreversible and can assess immune regulation. \u003C\u002Fp>\n\u003Ch3>4.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[4-1] Tabebordbar M, et al. Directed evolution of muscle-directed AAV capsids across species. Cell. 2021;184:4919-4938.e22. [H]\u003C\u002Fp>\n\u003Cp>[4-2] Weinmann J, et al. Identification of a myotropic AAV by barcoded in vivo evaluation. Nat Commun. 2020;11:5432. DOI:10.1038\u002Fs41467-020-19230-w. PMID:33116134. [H]\u003C\u002Fp>\n\u003Cp>[4-3] Gushchina LV. Intramuscular and intravenous AAV delivery in mouse models. Methods Mol Biol. 2026;2975:251-261. [N]\u003C\u002Fp>\n\u003Cp>[4-4] Intramuscular injection of AAV8 in mice and macaques is associated with substantial hepatic targeting and transgene expression. Gene Ther. 2014. PMID:25393537. [H]\u003C\u002Fp>\n\u003Cp>[4-5] Kornegay JN, et al. Assessment of systemic AAV-microdystrophin gene therapy in the GRMD model of Duchenne muscular dystrophy. Sci Transl Med. 2023;15(679):eabo1815. DOI:10.1126\u002Fscitranslmed.abo1815. [N]\u003C\u002Fp>\n\u003Cp>[4-6] Bridging from intramuscular to limb perfusion delivery of rAAV: optimization in a non-human primate study. Mol Ther Methods Clin Dev. 2019. PMID:30828586. [H]\u003C\u002Fp>","2026-09-18 11:31:54","2026-10-19 11:07:22",{"id":293,"service_pic":7,"service_title":294,"service_class":268,"class_name":269,"service_content":295,"service_tag":271,"created_at":296,"updated_at":297},51,"Skeletal Tissue","\u003Ch3>5.1 Bone tissue structure\u003C\u002Fh3>\n\u003Cp>Bone, bone marrow, and cartilage have different blood supplies, matrix, and cell turnover rates; osteoblasts, osteocytes, chondrocytes, or bone marrow cells should be designated first. \u003C\u002Fp>\n\u003Ch3>5.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>Native AAV9\u002FAAV-DJ can be used as a starting point, and systemic bone targeting usually requires capsid or ligand optimization. \u003C\u002Fp>\n\u003Ch3>5.3 Bone-specific promoter\u003C\u002Fh3>\n\u003Cp>COL1A1 is biased toward the osteogenic lineage, BGLAP is biased toward mature osteoblasts, and COL2A1 is biased toward chondrocytes. \u003C\u002Fp>\n\u003Ch3>5.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Intra-articular injection (IA): Targets the synovium. For mice, 0.5×10^10-1×10^11 vg\u002Fjoint, 5 μL, 30 G Hamilton, insert the needle at the medial edge of the patellar tendon of the knee joint; for rats, 1×10^10-2.5×10^10 vg\u002Fjoint, 50 μL; expression can last for >6 months [5-4,5-5]. \u003C\u002Fp>\n\u003Cp>Local\u002Fsystemic administration: 1×10^10-1×10^11 vg for local fracture; 1×10^11-1×10^12 vg\u002Ffor system. The joint cavity mainly transduces synovial lining cells, and the cartilage matrix barrier restricts AAV from penetrating cartilage and reaching chondrocytes. Treatment of cartilage lesions requires direct intrachondral injection or optimization of capsid engineering [5-5]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: Mainly synovial fibroblasts, with very limited chondrocyte transduction; intra-articular injection of AAV2.5-IL-1Ra in the equine OA model achieves sustained intra-articular IL-1Ra expression, no structural progression, and reduced lameness [5-6]. Bone tissue sampling and decalcification will affect nucleic acid, protein and fluorescence readout, and the detection method needs to be matched in advance. \u003C\u002Fp>\n\u003Ch3>5.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Xing et al. confirmed that intra-articular AAV-PRG4 gene therapy prevents post-traumatic osteoarthritis [5-4]; Frisch et al. reported the persistence and localization of transgene expression after a single AAV joint injection [5-5]; AAV2.5-IL-1Ra achieved safe and sustained intra-articular expression in the human OA knee joint in Phase I clinical trials [5-6]. \u003C\u002Fp>\n\u003Ch3>5.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[5-1] Yang YS, et al. Bone-targeting AAV silencing of Schnurri-3 prevents bone loss. Nat Commun. 2019;10:2958. [H]\u003C\u002Fp>\n\u003Cp>[5-2] Lee LR, et al. AAV vectors for local fracture and systemic skeleton delivery. Mol Ther Methods Clin Dev. 2019;15:101-111. [H]\u003C\u002Fp>\n\u003Cp>[5-3] Yoon DS, et al. AAV serotype selectivity for chondrocytes and cartilage. Int J Med Sci. 2021;18:3353-3360. [H]\u003C\u002Fp>\n\u003Cp>[5-4] Xing Y, et al. Intra-articular AAV-mediated PRG4 gene therapy for preventing posttraumatic osteoarthritis. Mol Ther. 2022;30:1864-1876. DOI:10.1016\u002Fj.ymthe.2022.02.015. [N]\u003C\u002Fp>\n\u003Cp>[5-5] Frisch J, et al. Persistence, localization, and external control of transgene expression after single injection of AAV into injured joints. Hum Gene Ther. 2013;24:423-433. DOI:10.1089\u002Fhum.2012.118. [H]\u003C\u002Fp>\n\u003Cp>[5-6] A phase 1 clinical trial shows safe, sustained AAV-mediated expression of IL-1Ra in human OA knee. Sci Transl Med. 2025. DOI:10.1126\u002Fscitranslmed.adu9804. [N]\u003C\u002Fp>","2026-09-18 11:32:14","2026-10-23 18:02:10",{"id":299,"service_pic":7,"service_title":300,"service_class":268,"class_name":269,"service_content":301,"service_tag":271,"created_at":302,"updated_at":303},52,"Lung","\u003Ch3>6.1 Lung tissue structure\u003C\u002Fh3>\n\u003Cp>Accessibility of airway epithelium, alveolar type I\u002FII cells, endothelium, and immune cells is affected by the distribution of mucus, barriers, and drug deposits. \u003C\u002Fp>\n\u003Ch3>6.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>Both AAV5\u002F6\u002F9 and lung screening capsids can be evaluated; in vitro airway models are not a substitute for in vivo deposition validation. \u003C\u002Fp>\n\u003Ch3>6.3 Lung-specific promoter\u003C\u002Fh3>\n\u003Cp>SFTPC is used for alveolar type II cells, SCGB1A1 is used for club cells, and FOXJ1 is used for ciliated cells. \u003C\u002Fp>\n\u003Ch3>6.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Intratracheal instillation (i.t.): Targeting the lower lung parenchyma. Mice 4×10^11-1×10^12 vg\u002Fmouse, 25-50 μL, video laryngoscope + Penn-Century nebulizer or 31 G tracheostomy needle; rats 1×10^11-5×10^12 vg\u002Fmouse, 100-300 μL, 22 G-ball head gavage needle; mainly transduces AT2, AT1 and bronchial epithelium, with more deposition in the lower lobe of the right lung, and peak expression in 1-2 weeks [6-4,6-5]. \u003C\u002Fp>\n\u003Cp>Intranasal instillation (i.n.): full respiratory tract coverage. Mice were divided into 25-50 μL into both nostrils and kept in a supine position for 1 minute; the upper respiratory tract (nasal mucosa) was additionally transduced, and there was much residue in the nasal cavity [6-4]. \u003C\u002Fp>\n\u003Cp>Aerosol inhalation: close to physiological exposure. NHP atomization is about 3 mL\u002Fanimal (Aerogen Solo vibrating mesh), general anesthesia + mechanical ventilation is about 30 minutes; sheep 10 mL\u002Ftime via mask or intubation, repeated for 9 months, has good tolerance; distribution is more uniform but the efficiency is only 10-30%. High doses (>2×10^12 vg) can penetrate the alveolar-capillary barrier and enter the blood, causing liver expression [6-4,6-5]. \u003C\u002Fp>\n\u003Ch3>6.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Akahori et al. established a non-invasive AAV airway pathway to target lung parenchymal cells and constructed a SARS-CoV-2 susceptible mouse model [6-4]; Wootton et al. systematically compared the AAV distribution in the mouse respiratory tract of four administration methods [6-5]; Körbelin et al. achieved lung targeting through NGS-guided capsid library screening [6-1]; Kang et al. lung-tropic AAV improved the survival of surfactant protein B-deficient mice [6-2]. The atomization\u002Fdrip method, inhalation efficiency, lobar distribution and liver off-target should be recorded. \u003C\u002Fp>\n\u003Ch3>6.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[6-1] Körbelin J, et al. Pulmonary targeting by NGS-guided AAV capsid library screening. Mol Ther. 2016;24:1050-1061. DOI:10.1038\u002Fmt.2016.62. [H]\u003C\u002Fp>\n\u003Cp>[6-2] Kang MH, et al. A lung-tropic AAV vector in surfactant B deficiency. Nat Commun. 2020;11:3929. [H]\u003C\u002Fp>\n\u003Cp>[6-3] Wu H, et al. Tropism of AAV serotypes in mouse lungs via intratracheal instillation. Virol J. 2024;21:302. [N]\u003C\u002Fp>\n\u003Cp>[6-4] Akahori Y, et al. Non-invasive administration of AAV to target lung parenchymal cells and develop SARS-CoV-2-susceptible mice. Commun Biol. 2022;5:45. DOI:10.1038\u002Fs42003-021-02984-3. [N]\u003C\u002Fp>\n\u003Cp>[6-5] Wootton SK, et al. AAV vector distribution in the mouse respiratory tract following four different methods of administration. BMC Biotechnol. 2017;17:43. DOI:10.1186\u002Fs12896-017-0365-2. [H]\u003C\u002Fp>","2026-09-18 11:32:33","2026-10-20 07:32:20",{"id":305,"service_pic":7,"service_title":306,"service_class":268,"class_name":269,"service_content":307,"service_tag":271,"created_at":308,"updated_at":309},53,"Kidney","\u003Ch3>7.1 Kidney tissue structure\u003C\u002Fh3>\n\u003Cp>Barrier differences in glomeruli, proximal\u002Fdistal tubules, collecting ducts, and interstitial cells make the results of systemic administration highly heterogeneous. \u003C\u002Fp>\n\u003Ch3>7.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>The effects of AAV9 and AAV2\u002F8 vary depending on the route and cells; local renal vein or retrograde ureter is often better than simple systemic exposure. \u003C\u002Fp>\n\u003Ch3>7.3 Kidney-specific promoter\u003C\u002Fh3>\n\u003Cp>NPHS1\u002F2 is used in podocytes, CDH16 is used in renal tubules, and AQP2 is used in collecting duct-related design. \u003C\u002Fp>\n\u003Ch3>7.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Renal vein injection: one of the best routes for kidney targeting. Mouse 1×10^11-1×10^12 vg\u002Fkidney (about half of the tail vein dose), 100 μL, 30-31 G needle, open the abdomen and microscopically clamp the renal artery and vein, inject slowly through the proximal end of the renal vein, remove the needle for compression after about 5 seconds; transduce the renal tubular epithelium and glomerular cells, the liver off-target is significantly lower than the tail vein, and detected in 2-4 weeks [7-3]. \u003C\u002Fp>\n\u003Cp>Retrograde ureteral injection: full nephron segment coverage. Mouse 1×10^11 vg\u002Fkidney, about 100 μL, puncture the ureter with a 33 G needle, clamp the renal artery for 30 minutes to maximize exposure, and inject slowly retrogradely at about 50-100 μL\u002Fmin; transduce proximal tubules, distal tubules, collecting ducts, and podocytes, and cooperate with segment-specific promoters such as SGLT2\u002FNKCC2\u002FECAD to achieve segment-specific expression; extrarenal expression is minimal [7-1]. \u003C\u002Fp>\n\u003Cp>Systemic administration: The renal transduction efficiency is low, 1×10^11-1×10^12 vg\u002Fanimal. Rat 1×10^11-5×10^11 vg\u002Fkidney, 100-200 μL; NHP data limited. Renal tissue vg, target cell expression, and urine\u002Fblood function indicators must be interpreted separately [7-4]. \u003C\u002Fp>\n\u003Ch3>7.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Rocca et al. comparatively confirmed that rAAV9 combined with renal vein injection is the optimal solution for kidney-targeted gene delivery [7-3]; Asico et al. achieved nephron segment-specific AAV gene expression [7-1]; Lee et al. compared the renal gene delivery efficiency of adenovirus\u002FAAV\u002Flentivirus via intravenous and direct renal injection [7-4]. \u003C\u002Fp>\n\u003Ch3>7.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[7-1] Asico LD, et al. Nephron segment-specific gene expression using AAV vectors. Biochem Biophys Res Commun. 2018;497:19-24. [H]\u003C\u002Fp>\n\u003Cp>[7-2] Lake BB, et al. An atlas of healthy and injured cell states and niches in the human kidney. Nature. 2023. [N\u002FH]\u003C\u002Fp>\n\u003Cp>[7-3] Rocca CJ, et al. rAAV9 with renal vein injection for kidney-targeted delivery. Gene Ther. 2014;21:618-628. [F]\u003C\u002Fp>\n\u003Cp>[7-4] Lee PY, et al. Comparison of gene delivery to the kidney by adenovirus, AAV, and lentiviral vectors after intravenous and direct kidney injections. Hum Gene Ther. 2020;31:463-477. PMC6919283. [H]\u003C\u002Fp>","2026-09-18 11:32:56","2026-10-23 19:17:10",{"id":311,"service_pic":7,"service_title":312,"service_class":268,"class_name":269,"service_content":313,"service_tag":271,"created_at":314,"updated_at":315},54,"Pancreas","\u003Ch3>8.1 Pancreatic tissue structure\u003C\u002Fh3>\n\u003Cp>Islet cells are spatially adjacent to exocrine and ductal cells, and expression in tissue homogenates can easily mask cell-type off-targeting. \u003C\u002Fp>\n\u003Ch3>8.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV6\u002F8\u002F9 can be used as comparison objects; retrograde administration through the pancreatic duct often increases local exposure, but the technical requirements are higher. \u003C\u002Fp>\n\u003Ch3>8.3 Pancreas-specific promoter\u003C\u002Fh3>\n\u003Cp>INS1\u002F2 is used for β cells, GCG is used for α cells, and PDX1 needs to be interpreted in conjunction with development and disease states. \u003C\u002Fp>\n\u003Ch3>8.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Retrograde perfusion of the pancreatic duct: targeting the exocrine part of the pancreas. Mice 2×10^11-2×10^12 vg\u002Fmouse (AAV6), 100 μL, intubated through duodenal papilla\u002Fcommon bile duct, peristaltic pump 10 μL\u002Fmin×10 min perfusion; the proximal hepatic end of the common bile duct is clamped to prevent viruses from entering the liver; AAV6 has the highest transduction of acinar cells (about 37% acinar expression), ductal cells are also transduced, and islet cell transduction is limited; the Ela promoter is acinar-specific and can be detected in 4-6 weeks [8-1,8-5]. \u003C\u002Fp>\n\u003Cp>Systemic administration: 1×10^11-1×10^12 vg\u002Fanimal. It is important to note that the liver and acinar cells are off-target. Acute pancreatitis can be induced if the perfusion rate is too fast (>50 μL\u002Fmin) or the pressure is too high, and the peristaltic pump must be accurately controlled [8-4]. \u003C\u002Fp>\n\u003Ch3>8.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Quirin et al. established an AAV retrograde pancreatic duct gene delivery protocol [8-1]; Orabi et al. targeted inhibition of calcineurin in pancreatic acinar cells to prevent post-ERCP pancreatitis [8-4]; Xiao et al. established a Nat Protoc protocol for pancreatic duct infusion of AAV and cell labeling dyes for pancreatic cell lineage tracing and genetic manipulation [8-5]. It is recommended to verify islet co-staining, hormone secretion and blood glucose phenotypes. \u003C\u002Fp>\n\u003Ch3>8.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[8-1] Quirin KA, et al. AAV retrograde pancreatic ductal gene delivery. Mol Ther Methods Clin Dev. 2018;8:8-20. [H]\u003C\u002Fp>\n\u003Cp>[8-2] Chen M, et al. Capsid-optimized AAV8 delivery to pancreas and pancreatic tumors. Hum Gene Ther Methods. 2017;28:49. [H]\u003C\u002Fp>\n\u003Cp>[8-3] Ramzy A, et al. AAV8 Ins1-Cre recombination and off-target considerations. Sci Rep. 2020;10:10518. [H]\u003C\u002Fp>\n\u003Cp>[8-4] Orabi AI, et al. Targeted inhibition of pancreatic acinar cell calcineurin prevents post-ERCP pancreatitis. Cell Mol Gastroenterol Hepatol. 2017;4:247-262. DOI:10.1016\u002Fj.jcmgh.2017.04.008. [H]\u003C\u002Fp>\n\u003Cp>[8-5] Xiao X, Guo P, Prasadan K. Pancreatic cell tracing, lineage tagging and targeted genetic manipulations in multiple cell types using pancreatic ductal infusion of AAV vectors and\u002For cell-tagging dyes. Nat Protoc. 2014;9:2719-2724. DOI:10.1038\u002Fnprot.2014.183. [F]\u003C\u002Fp>","2026-09-18 11:33:13","2026-10-19 10:56:40",{"id":317,"service_pic":7,"service_title":318,"service_class":268,"class_name":269,"service_content":319,"service_tag":271,"created_at":320,"updated_at":321},55,"Adipose Tissue","\u003Ch3>9.1 Adipose tissue structure\u003C\u002Fh3>\n\u003Cp>White, brown, and beige fat differ in blood supply, cellularity, and promoter activity; mature adipocytes must be distinguished from the stromal vascular component. \u003C\u002Fp>\n\u003Ch3>9.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV8\u002F9 is a common starting point. If necessary, use fat to screen capsids; systemic administration is prone to liver dominance. \u003C\u002Fp>\n\u003Ch3>9.3 Fat-specific promoter\u003C\u002Fh3>\n\u003Cp>ADIPOQ favors mature adipocytes, UCP1 favors brown\u002Fbeige fat, and FABP4 may be expressed in other cells. \u003C\u002Fp>\n\u003Ch3>9.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Local injection into the fat pad: targeting white\u002Fbrown fat. Mouse 1×10^11-2×10^11 vg\u002Fpad (AAV8), eWAT 30-50 μL\u002Fpad divided into 2 points, iBAT 20 μL\u002Fleaf (a total of 40 μL for two leaves), iWAT 25 μL\u002Fside; 30 G Hamilton (use 32 for BAT G fine injection), isoflurane anesthesia; AAV8 is the most efficient in transducing adipose tissue, the ADIPOQ promoter is specific for mature adipocytes, and the expression lasts for >4 weeks [9-2,9-5]. \u003C\u002Fp>\n\u003Cp>Subcutaneous\u002Fsystemic administration: SC fat pad local regimen 2×10^10 vg\u002Fside, 25-30 μL\u002Fpoint; system 1×10^11-1×10^12 vg\u002Fanimal. Brown fat (BAT) dosage is approximately 1×10^9-2×10^10 vg\u002Fpad. Each point of eWAT is ≤10 μL to prevent leakage into the abdominal cavity [9-4]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: mature adipocytes + stromal vascular component; off-targets include intraperitoneal leakage (eWAT) and high-dose hepatic spread. Adipocyte expression, liver off-target and metabolic phenotypes should be detected simultaneously; diet and temperature are important confounding factors [9-1]. \u003C\u002Fp>\n\u003Ch3>9.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Jimenez et al. achieved in vivo AAV genetic engineering of white and brown adipose tissue in adult mice [9-2]; Bates et al. reviewed adipose tissue as an emerging target of AAV [9-1]; research on subcutaneous fat tropism gene therapy for congenital leptin defects demonstrates the SC pathway [9-4]. \u003C\u002Fp>\n\u003Ch3>9.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[9-1] Bates R, Huang W, Cao L. Adipose tissue as an emerging target for AAV vectors. Mol Ther Methods Clin Dev. 2020;19:236-249. [H]\u003C\u002Fp>\n\u003Cp>[9-2] Jimenez V, et al. AAV-mediated genetic engineering of white and brown adipose tissue. Diabetes. 2013;62:4012-4022. [F]\u003C\u002Fp>\n\u003Cp>[9-3] Sancar G, et al. FGF1 and insulin control lipolysis by convergent pathways. Cell Metab. 2022;34:171-183.e6. [H]\u003C\u002Fp>\n\u003Cp>[9-4] Subcutaneous administration of adipose-tropic gene therapy for congenital leptin deficiency. 2025. PMC12288548. [N]\u003C\u002Fp>\n\u003Cp>[9-5] Direct aden-associated virus injection of murine adipose tissue. Methods Mol Biol. 2023. PMC10213077. [F]\u003C\u002Fp>","2026-09-18 11:33:32","2026-10-18 10:24:01",{"id":323,"service_pic":7,"service_title":324,"service_class":268,"class_name":269,"service_content":325,"service_tag":271,"created_at":326,"updated_at":327},56,"Skin","\u003Ch3>10.1 Skin tissue structure\u003C\u002Fh3>\n\u003Cp>Keratinocytes, fibroblasts, hair follicles and immune cells are clearly stratified, and the stratum corneum forms an important delivery barrier. \u003C\u002Fp>\n\u003Ch3>10.2 AAV serotype selection\u003C\u002Fh3>\n\u003Cp>AAV2\u002F6\u002F8\u002F9 can be compared by cell and wound status; in vitro keratinocyte results need to be reviewed in intact skin or wounds. \u003C\u002Fp>\n\u003Ch3>10.3 Skin-specific promoter\u003C\u002Fh3>\n\u003Cp>KRT14\u002FKRT5 favors basal keratinocytes, IVL favors differentiated keratinocytes, and COL1A1 favors fibroblasts. \u003C\u002Fp>\n\u003Ch3>10.4 Administration method and dosage reference\u003C\u002Fh3>\n\u003Cp>Intradermal injection: targeting the basal layer of the epidermis and dermis. Mouse working concentration is 1×10^12 vg\u002FmL, approximately 2×10^10 vg per point, 10-20 μL per point, 29-31 The tip of the G insulin needle is inserted into the skin at an angle of 10-15°, and the wheal is confirmed to be in the skin; AAV2\u002F6 transduces basal keratinocytes (KRT14 promoter), and COL1A1 targets dermal fibroblasts; it is expressed for 2-4 weeks [10-3]. \u003C\u002Fp>\n\u003Cp>Local application on wounds: intradermally inject a total of 80-120 μL at 4-6 points per wound, or drop it directly onto the wound bed and then cover it with Vaseline gauze\u002Fhydrogel scaffold to prevent loss; wound repair cells (keratinocytes, fibroblasts, macrophages) can all be transduced. 50-100 μL per point in rats; the structure of pig skin is highly similar to that of humans, making it a transformation model [10-1, 10-3]. \u003C\u002Fp>\n\u003Cp>Coverage and limitations: The intact stratum corneum of skin is the main barrier for transdermal penetration, and intradermal injection is limited to a few millimeters around the injection point; injection depth, positive area, cell co-localization, inflammation and wound repair endpoints should be reported [10-2]. \u003C\u002Fp>\n\u003Ch3>10.5 Key research evidence\u003C\u002Fh3>\n\u003Cp>Keswani et al. reported the tropism of AAV vectors in wound healing in mice [10-3]; Kurita et al. generated skin epithelial tissue through in vivo reprogramming of wound-resident cells [10-1]; Shen et al. optimized AAV for in vitro and in vivo delivery of epidermal keratinocytes [10-2]. \u003C\u002Fp>\n\u003Ch3>10.6 References for this chapter\u003C\u002Fh3>\n\u003Cp>[10-1] Kurita M, et al. In vivo reprogramming of wound-resident cells generates skin epithelial tissue. Nature. 2018;561:243-247. [H]\u003C\u002Fp>\n\u003Cp>[10-2] Shen Q, et al. Optimization of an AAV vector for epidermal keratinocytes in vitro and in vivo. J Dermatol Sci. 2024;115:101-110. [N]\u003C\u002Fp>\n\u003Cp>[10-3] Keswani SG, et al. AAV vector tropism in murine wound healing. Wound Repair Regen. 2012;20:592-600. [F]\u003C\u002Fp>","2026-09-18 11:33:56","2026-10-19 13:39:21",{"id":329,"service_pic":7,"service_title":330,"service_class":6,"class_name":9,"service_content":331,"service_tag":11,"created_at":332,"updated_at":333},57,"Special Topics in Neuroscience","\u003Ch2>\u003Ca name=\"_Toc240028057\">\u003C\u002Fa>Determine the type of research first\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"120\">\n\u003Cp>\u003Cstrong>Type\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"264\">\n\u003Cp>\u003Cstrong>Priority\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>\u003Cstrong>Common Misunderstandings\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"120\">\n\u003Cp>Local expression\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"264\">\n\u003Cp>Brain region, cell type, diffusion radius, needle track control\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Extrapolate local high expression to whole-brain ability\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"120\">\n\u003Cp>Retrograde tracing\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"264\">\n\u003Cp>Accessibility of projection terminals, retrograde capsid, direction and synaptic specificity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Equating retrograde uptake across synapses\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"120\">\n\u003Cp>Whole-brain\u002Fwidely expressed\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"264\">\n\u003Cp>Species\u002Fstrain, BBB receptors, intravenous dose and hepatic off-target\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Directly copy the PHP capsid\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"120\">\n\u003Cp>CSF delivery\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"264\">\n\u003Cp>Intrathecal\u002Fcisternal\u002Fventricular pathways, meninges and DRG exposure\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"211\">\n\u003Cp>Only look at brain tissue fluorescence and ignore neurotoxicity\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028058\">\u003C\u002Fa>Situations when neural projects must be upgraded\u003C\u002Fh2>\n\u003Cul>\n\u003Cli>Engineering capsids across the blood-brain barrier or NHP project. \u003C\u002Fli>\n\u003Cli>Study on the safety of CSF high dose, long-term expression or sensory nerves. \u003C\u002Fli>\n\u003Cli>Transsynaptic tools, cytotoxic effectors, long-term calcium\u002Fvoltage sensor expression. \u003C\u002Fli>\n\u003Cli>Projects that require cross-strain, cross-species replication or clinical transformation. \u003C\u002Fli>\n\u003C\u002Ful>\n\u003Cp>Recent evidence shows that receptor reprogramming can significantly improve brain delivery, but this advantage is tied to receptor expression and model selection; neurological projects should consider brain area coverage, cell types, liver\u002Fperipheral off-targeting and DRG safety as concurrent endpoints. [15-3 to 15-5]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028059\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[15-1] Tervo DGR, et al. A designer AAV variant permits efficient retrograde access to projection neurons. Neuron. 2016;92:372-382. [H]\u003C\u002Fp>\n\u003Cp>[15-2] Deverman BE, et al. Cre-dependent selection yields AAV variants for widespread adult brain gene transfer. Nat Biotechnol. 2016;34:204-209. DOI:10.1038\u002Fnbt.3440. [H]\u003C\u002Fp>\n\u003Cp>[15-3] Goertsen D, et al. Brain-wide AAV capsid variants with decreased liver targeting in mouse and marmoset. Nat Neurosci. 2022;25:106-115. DOI:10.1038\u002Fs41593-021-00969-4. [H]\u003C\u002Fp>\n\u003Cp>[15-4] Ling Q, et al. AAV-based in vivo gene therapy for neurological disorders. Nat Rev Drug Discov. 2023;22:789-806. DOI:10.1038\u002Fs41573-023-00766-7. PMID:37658167. [N\u002FH]\u003C\u002Fp>\n\u003Cp>[15-5] Huang Q, et al. Human transferrin receptor-binding AAV for brain-wide delivery. Science. 2024;384:1220-1227. DOI:10.1126\u002Fscience.adm8386. [N]\u003C\u002Fp>","2026-09-18 11:34:19","2026-10-20 01:14:24",{"id":335,"service_pic":7,"service_title":336,"service_class":6,"class_name":9,"service_content":337,"service_tag":11,"created_at":338,"updated_at":339},58,"Tissue-Application Evidence Grading","\u003Cp>Evidence ratings are used to describe \"closeness to the current project\" and are not a substitute for journal impact factors. High-impact literature can increase credibility, but only if the species, strain, capsid, expression cassette, route of administration, dose, and endpoint are close enough to serve as a direct basis for design. \u003C\u002Fp>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"62\">\n\u003Cp>\u003Cstrong>Level\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"331\">\n\u003Cp>\u003Cstrong>Definition\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>\u003Cstrong>Recommended usage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"62\">\n\u003Cp>A\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"331\">\n\u003Cp>Same species\u002Fstrain, same tissue cells, same pathway, and have quantitative and functional endpoints\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>Can be used as the core basis for the main plan and dosage interval\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"62\">\n\u003Cp>B\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"331\">\n\u003Cp>Same species and target tissue, but one difference in capsid, expression cassette or pathway\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>Supports candidate sorting, still needs pre-experimentation\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"62\">\n\u003Cp>C\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"331\">\n\u003Cp>Different species or similar only at the organizational level\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>Used to propose hypotheses, not to promise to reproduce\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"62\">\n\u003Cp>D\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"331\">\n\u003Cp>In vitro, single reporter gene, supplier information or no complete method\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"202\">\n\u003Cp>For clues only, independent verification is required\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>Clinical success demonstrates the feasibility of the AAV platform, but it cannot replace the dosage and safety verification of research-grade vectors in specific models. High-quality clinical studies of the eye, liver, and muscle\u002Fnervous system can be used in the risk framework; specific experiments should still be based on evidence from the same model. [16-1 to 16-4]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028061\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[16-1] Mendell JR, et al. Single-dose gene-replacement therapy for spinal muscular atrophy. N Engl J Med. 2017;377:1713-1722. DOI:10.1056\u002FNEJMoa1706198. PMID:29091557. [H]\u003C\u002Fp>\n\u003Cp>[16-2] Russell S, et al. Voretigene neparvovec phase 3 trial. Lancet. 2017;390:849-860. DOI:10.1016\u002FS0140-6736(17)31868-8. [H]\u003C\u002Fp>\n\u003Cp>[16-3] Pipe SW, et al. Gene therapy with etranacogene dezaparvovec for hemophilia B. N Engl J Med. 2023;388:706-718. DOI:10.1056\u002FNEJMoa2211644. [N\u002FH]\u003C\u002Fp>\n\u003Cp>[16-4] Lv J, et al. AAV1-hOTOF gene therapy for autosomal recessive deafness 9. Lancet. 2024;403:2317-2325. DOI:10.1016\u002FS0140-6736(23)02874-X. PMID:38280389. [N]\u003C\u002Fp>","2026-09-18 11:34:38","2026-10-29 00:29:29",{"id":341,"service_pic":7,"service_title":342,"service_class":6,"class_name":9,"service_content":343,"service_tag":11,"created_at":344,"updated_at":345},59,"Natural serotypes, engineered capsids and screening","\u003Cp>Native serotypes are suitable for projects with well-documented, cycle-limited and acceptable tissue distribution; engineered capsids are used to address receptor restriction, tissue barriers, immune evasion or off-target issues. Engineering strategies include rational design, random peptide insertion, DNA recombination, ancestry reconstruction, and machine learning design. Performance evaluation must be broken down into assembly, packaging, stabilization, receptor binding, tissue entry, cellular expression, and immune response. \u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028063\">\u003C\u002Fa>Full screening workflow\u003C\u002Fh2>\n\u003Ctable>\n\u003Cthead>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>\u003Cstrong>Stage\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>\u003Cstrong>Core Movements\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>\u003Cstrong>Main phase-out risks\u003C\u002Fstrong>\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Fthead>\n\u003Ctbody>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>1 Target product portrait\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Define species, cells, pathways, doses, off-targets and production indicators\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Blurred targets lead to drift in the screening direction\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>2 Library design\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Control insertion sites, mutation depth, theoretical diversity and manufacturability\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>A large number of unassembleable sequences\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>3 Production and input library quality control\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Sequencing input library, titer, assembly and packaging bias\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Mistaking production advantages for organizational advantages\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>4 In vivo selection\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Biological replicates, negative organs, multiple doses\u002Ftime points\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Bottlenecks, random drift and off-target enrichment\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>5 NGS and models\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Normalize by input library to differentiate between DNA arrival, RNA expression and function\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Label swapping, PCR bias and overfitting\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>6 Monoclonal retest\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Independent preparation, blind comparison, histology and functional verification\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Pool filtering results cannot be reproduced\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003Ctr>\n\u003Ctd width=\"106\">\n\u003Cp>7 Conversion Review\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"312\">\n\u003Cp>Cross-strain\u002Fspecies, production scale-up, immunity and safety\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003Ctd width=\"178\">\n\u003Cp>Early performance cannot be migrated across systems\u003C\u002Fp>\n\u003C\u002Ftd>\n\u003C\u002Ftr>\n\u003C\u002Ftbody>\n\u003C\u002Ftable>\n\u003Cp> \u003C\u002Fp>\n\u003Cp>High-throughput sequencing and machine learning can expand the search space, but are not a substitute for rigorous input library calibration and independent validation. Reviews in the past three years further emphasized that incorporating assembly fitness, tissue performance and production attributes into multi-objective optimization is superior to ranking only by a single enrichment fold. [17-3 to 17-5]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028064\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[17-1] Deverman BE, et al. Cre-dependent selection yields AAV variants for widespread adult brain gene transfer. Nat Biotechnol. 2016;34:204-209. [H]\u003C\u002Fp>\n\u003Cp>[17-2] Ogden PJ, et al. Comprehensive AAV capsid fitness landscape enables machine-guided design. Science. 2019;366:1139-1143. DOI:10.1126\u002Fscience.aaw2900. PMID:31780559. [H]\u003C\u002Fp>\n\u003Cp>[17-3] Bryant DH, et al. Deep diversification of an AAV capsid protein by machine learning. Nat Biotechnol. 2021;39:691-696. DOI:10.1038\u002Fs41587-020-00793-4. PMID:33574611. [H]\u003C\u002Fp>\n\u003Cp>[17-4] Guo J, et al. Computationally guided AAV engineering for enhanced gene delivery. Trends Biochem Sci. 2024;49:457-469. DOI:10.1016\u002Fj.tibs.2024.03.002. PMID:38531696. [N]\u003C\u002Fp>\n\u003Cp>[17-5] Tan F, et al. Artificial intelligence-based approaches for AAV vector engineering. Adv Sci. 2025;12:e2411062. DOI:10.1002\u002Fadvs.202411062. PMID:39932449. [N]\u003C\u002Fp>","2026-09-18 11:34:57","2026-10-19 10:25:14",{"id":347,"service_pic":7,"service_title":348,"service_class":6,"class_name":9,"service_content":349,"service_tag":11,"created_at":350,"updated_at":351},60,"Translational Risks in Capsid Development","\u003Ch2>\u003Ca name=\"_Toc240028066\">\u003C\u002Fa>Five types of conversion risks\u003C\u002Fh2>\n\u003Ch3>Species barrier\u003C\u002Fh3>\n\u003Cp>Differences in receptors, glycans, endothelial barriers, and immune background can cause mouse enrichment results to disappear in NHP or human systems. The solution is to add human cells\u002Forganoids, second lines and cross-species validation as early as possible. \u003C\u002Fp>\n\u003Ch3>Production adaptation\u003C\u002Fh3>\n\u003Cp>Highly transducing candidates may be poorly assembled, have low titers, have many empty shells, or be unstable in the scale-up process. Yield, VP ratio, genome integrity and freeze-thaw stability thresholds should be set in the early stage of screening. \u003C\u002Fp>\n\u003Ch3>Library bias\u003C\u002Fh3>\n\u003Cp>Systematic deviations exist between plasmid libraries, virus input libraries, and tissue recovery libraries. Input libraries must be sequenced and bottlenecks estimated with biological replicates and negative tissue. \u003C\u002Fp>\n\u003Ch3>Off-target enrichment\u003C\u002Fh3>\n\u003Cp>High exposure to liver, spleen or endothelium can produce strong enrichment masking target cells. RNA expression, cell sorting, and non-target organs should be analyzed in parallel. \u003C\u002Fp>\n\u003Ch3>Intellectual Property\u003C\u002Fh3>\n\u003Cp>Sequences, insertion sites, screening methods and uses may be individually protected; sequence searches and free implementation evaluations should be completed before candidates enter amplification. \u003C\u002Fp>\n\u003Cp>Safety risks are not a simple ranking of \"low natural capsid, high engineered capsid\". Dosage, receptor distribution, expression cassette toxicity, impurities, and manufacturing processes all combine to modify risk. High-dose systemic administration and CSF administration particularly require validation that incorporates hepatotoxicity, complement\u002Fhematological events, DRG pathology, and non-target tissue expression. [18-2 to 18-5]\u003C\u002Fp>\n\u003Ch2>\u003Ca name=\"_Toc240028067\">\u003C\u002Fa>References for this chapter\u003C\u002Fh2>\n\u003Cp>[18-1] Hordeaux J, et al. Neurotropic properties of AAV-PHP.B are limited to C57BL\u002F6J mice. Mol Ther. 2018;26:664-668. [H]\u003C\u002Fp>\n\u003Cp>[18-2] Hinderer C, et al. Severe toxicity following high-dose intravenous AAV-SMN in NHP and piglets. Hum Gene Ther. 2018;29:285-298. DOI:10.1089\u002Fhum.2018.015. PMID:29378426. [H]\u003C\u002Fp>\n\u003Cp>[18-3] Hordeaux J, et al. AAV-induced dorsal root ganglion pathology. Hum Gene Ther. 2020;31:808-818. DOI:10.1089\u002Fhum.2020.167. [H]\u003C\u002Fp>\n\u003Cp>[18-4] Shen W, et al. rAAV immunogenicity, toxicity, and durability in 255 clinical trials. Front Immunol. 2022;13:1001263. [H]\u003C\u002Fp>\n\u003Cp>[18-5] Lek A, et al. Death after high-dose rAAV9 gene therapy in Duchenne muscular dystrophy. N Engl J Med. 2023;389:1203-1210. DOI:10.1056\u002FNEJMoa2307798. [N]\u003C\u002Fp>\n\u003Cp>[18-6] Park S, et al. DoE optimization of AAV2\u002FAAV9 production and full-capsid ratio. Biotechnol J. 2024;19:e2300667. DOI:10.1002\u002Fbiot.202300667. 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