Intravitreal AAV injections

Mice were anesthetized, and eyes dilated using 1% tropicamide as previously reported ¹⁶. AAV2 construct solution (final concentration, 1.8 × 10¹² GC/mL) was carefully administered (2 µl) through the sclera at a 45° into the vitreous towards the ora serrata and posterior to the temporal limbus avoiding contact with lens. Injection was performed using 33G needle connected to 5 µl Hamilton syringe (Hamilton AG) guided by a surgical microscope (Carl Zeiss) to facilitate accurate focusing and a period of 30 seconds was allowed before removing the needle to permit diffusion of the virus in order to prevent leakage from the injection track. Animals were monitored and sacrificed at specified time points as indicated.