AAV3 Luciferase-mCherry

Adeno-Associated Virus Serotype 3 (AAV3) shares 82% sequence homology with AAV2, and like AAV2, requires the Heparan Sulfate Proteoglycan (HSPG) receptor for cell attachment. AAV3 vectors transduce human liver cancer cells extremely efficiently because AAV3 utilizes the human Hepatocyte Growth Factor Receptor (hHGFR) as a co-receptor for viral entry, which is highly expressed in these cells. Both the extracellular and intracellular kinase domains of hHGFR are required for AAV3-mediated transgene expression. These AAV particles constitutively express the firefly (Photinus pyralis) luciferase and mCherry genes connected via a T2A linker, under the control of a CMV promoter. The T2A self-cleaving peptide (derived from Thosea asigna virus 2A) leads to the efficient cleavage of the transcript, and expression of luciferase and mCherry as two separate proteins.