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Applicable species: mice, rats, NHP (rhesus/cynomolgus), dogs, pigs [12-45, 12-62, 12-44]. Dose range: mouse 1×10^8-3×10^9 vg/eye; rat 1×10^9-1×10^10; NHP 7×10^11-7×10^12 vg/eye (AAV8, engineered capsid can be as low as 1×10^10); dog 1.0-1.4×10^11 vg/eye [12-45,12-62]. Injection volume: mice 1-2 μL; rats 2-5 μL; NHP 100-200 μL/eye (traditional transretinal approach); dogs 200 μL/eye [12-45,12-62]. Needle: Mouse 33 G Hamilton blunt needle or glass microelectrode; NHP 39 G injection cannula/36-42 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 (AAV8/5 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.