Available Vector Systems
Adenovirus Vectors:
The VCF is constructing and/or propagating recombinant adenoviral vectors for gene therapy applications. The adenoviral vector that is being used at the Facility is a serotype 5 first generation vector deleted for E1 and E3. The gene of interest is introduced into the adenovirus vector using a shuttle vector that carries a CMV expression cassette with a LoxP site. The shuttle plasmid is cotransfected into 293 cells expressing CRE recombinase (CRE8 cells) with the adenoviral backbone plasmid (psi 5) containing two LoxP sites in the E1 region. Recombination between the vector backbone and the shuttle plasmid is driven by the CRE recombinase, resulting in a high frequency of the appropriate recombinants. Thus, either the plaques are picked, expanded, and screened for expression of the transgene, or the viruses from the transfected CRE8 cells are pooled and passaged three to four times on CRE8 cells.