Renal vein injection, a surgery-based gene delivery method, has achieved exogenous gene expression in the kidney at different levels. In this protocol, we present the experimental procedure of the renal pelvis injection in the mouse as follows. Preparation
●Anaesthetize the mouse by subcutaneous injection of ketamine (100 mg/kg) and xylazine (10 mg/kg).
●Place it in a supine position and shave the left side of the back of the mouse from shoulder to rump and flank to spine with a shaver, and place vet ointment such as Oculentum simplex, Teva Pharmachemie in both eyes to prevent corneal desiccation.
●Prepare a 0.5 ml U-100 insulin syringe (29G needle) containing 100 µL virus to be injected for each anesthetized mouse, making sure there are no air bubbles.
Target coordinates and inject
●Surgically expose the left kidney of the mouse, then clamp the naked skin of the mouse with surgical clips. Use scissors to make an incision to the skin layer approximately 1 cm from the spine and below the ribcage.
●Locate the kidney and the adipose tissue around the kidneys, gently separate it from the surrounding fat just enough to visualize the renal pelvis, a small white dot.
●Inject the solution quickly as possible, or it will cause backflow and will greatly affect the efficiency of infection. Please note that avoid the renal vein, artery and ureter when injecting, and the depth of the insertion is about 0.5 cm.
Post-injection care
●Keep the needle in place for approximately 10 s to prevent backflow of the solution. Then carefully and slowly remove the needle. Return the organ to the abdominal cavity and suture the muscle layer of the animal with 5-0 absorbable sutures.
●Remove the mouse and place it prone on a heating pad for recovery.

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