Fig. 1.

Scalable PEI transfection of HEK293T cells produces secreted rAAV. a Schematic diagram of method for production of secreted rAAV. b Single-stranded (ss) rAAV2 vectors used in this study contained the cytomegalovirus (CMV) enhancer, hybrid chicken β-actin (hCBA) promoter, truncated CBA intron, rabbit beta-globin splice acceptor, woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), bovine growth hormone polyadenylation signal, and transgene encoding either red fluorescent protein (TurboRFP) or enhanced green fluorescent protein (EGFP). c Only co-transfection of both rAAV2/1 capsid helper (pDP.rs) and transgene (pAAV-hCBA-RFP) produces rAAV in the media capable of transducing PNGC. Bar, 100 μm d Genomic titers of all secreted rAAVs at 72 h post-transfection quantified as vector genomes/mL of culture media (Mean ± SEM N = 3)