AAV1-EF1a-GFP(FRT)

AAVs are attractive gene delivery vectors due to their ability to package and express foreign genes in a wide variety of tissues. Different AAV serotypes exhibit specific tissue tropisms and varying transduction efficiencies and have been shown to enable stable, long-term gene expression. They are able to infect both dividing and non-dividing cells without any known associated pathogenicity. Furthermore, recombinant AAV (rAAV) can be manufactured and purified at high titers, making them readily available for clinical use.

Two approaches have been used to manufacture rAAV gene therapy vectors. The first approach uses triple transfection, where cultured mammalian cells are transfected with three separate plasmids. One plasmid contains the therapeutic gene of interest flanked by viral ITRs, which are the only cis signals required for packaging into the AAV capsid. A separate plasmid encodes the rep and cap genes, while the final plasmid contains adenoviral helper genes essential for viral replication. The second approach uses the baculovirus/Sf9 system, where three separate viruses (Rep baculovirus, VP baculovirus, and AAV ITRs flanking the gene baculovirus) are used for infection. The resulting rAAV vector contains the transgene packaged into the selected rAAV capsid.