Recombinant Adeno Associated Virus Based Gene Delivery: A Technique for Transgene Delivery and Cardiac-Specific Expression in Rodent Model

After determining the viral titer according to the text protocol, prepare rAAV9 working solutions with virus titers of 1-7 x 10¹² particles per milliliter. To treat neonatal mice, use the rAAV9 solution to prefill a 0.33 x 12.7-milliliter insulin syringe with a 29-gauge 1/2-inch needle, avoiding air bubbles.

After cryoanesthetizing a pup, hold the pup in one hand with thumb and forefinger. Then, swipe the back skin of the pup with a swab stick saturated with 70% isopropyl alcohol. Insert the syringe needle into the anterior-dorsal subcutis of the animal at an angle of 5 to 10 degrees before injecting 50 to 70 microliters of the rAAV9 solution.