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Applicable species: mice, rats; NHP is a historical preclinical/clinical protocol (has been basically replaced by peripheral IV) [12-13]. Dose range: 1×10^10-1×10^12 vg/mouse (AAV8 can transduce 90-95% of hepatocytes through the portal vein); 1×10^11-1×10^12 vg/mouse [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 After the needle is withdrawn, press with a sterile cotton swab for 3-5 minutes to stop bleeding; the minimally invasive alternative is to inject 50 μL slowly through the spleen parenchyma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 (AAV8/9 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/thrombosis, non-repetitive administration [12-13].