Recombinant adenovirus and adeno-associated virus, cell lines and methods of production and use thereof
AssigneeThe Trustees of the University of Pennsylvania
InventorsJames M. Wilson, Krishna J. Fisher, Guang-Ping Gao
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Abstract
An adenovirus E1/E4 expressing packaging cell line is provided, which permits the generation of recombinant adenoviruses deleted in both gene regions. A method for enhancing the efficiency of transduction of a recombinant AAV into a target cell is provided by infecting a target cell with a recombinant AAV comprising a selected transgene under the control of regulatory sequences. The infected cell is contacted with an agent which facilitates the conversion of single stranded recombinant virus to its double stranded form.
Claims12
A method of enhancing conversion of single-stranded AAV to double-stranded AAV, said method comprising the step of culturing a host cell containing:
- (a) a nucleic acid molecule comprising AAV 5′ inverted terminal repeats (ITRs), a transgene under the control of regulatory sequences directing expression thereof, and AAV 3′ ITRs;
- (b) a nucleic acid molecule comprising a nucleic acid sequence encoding adenovirus E4 ORF6, wherein the E4 ORF6 is under the control of heterologous regulatory sequences directing expression thereof;
- (c) a nucleic acid molecule comprising a nucleic acid sequence encoding rep under the control of regulatory sequences directing expression thereof;
- (d) a nucleic acid molecule comprising nucleic acid sequence encoding cap under the control of regulatory sequences directing expression thereof.
Dependencies are inferred from the claim text.