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Applicable species: mice, rats, NHP, dogs [12-47]. Dose range: mice 1×10^9-1×10^10 vg/eye; rats 1×10^9-5×10^9; NHP 7×10^11 vg/eye (AAV8) [12-47]. Injection volume: 1-2 μL for mice; 5 μL for rats; NHP 100 μL/eye [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].