AAV-mediated gene expression in primary islets

Primary islets from wild-type mice or human donors were isolated for AAV treatment. 150 IEQ islets were treated with AAV2/9 viruses expressing CTCF or GFP control at the titer of 10¹² viral genomes per ml (vg/ml). Virus was administrated into cultured islets at the final concentration of 20 μl/ml culture medium. Islets were harvested at 5–7 d after transduction, enabling robust exogenous gene expression for Western blot analysis, calcium imaging, and perifusion assay.