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Applicable species: mice (subcutaneous/in 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/tumor; rats 1×10^10-5×10^10 vg/tumor [12-64]. Injection volume: mice 50-100 μL/tumor (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/system [12-64].