Patent ID: 6221646
Filing Date: 2001-04-24
Classification: C12N

Abstract:
A method for producing replication-defective recombinant AAV virions substantially free of wild-type AAV and helper adenovirus, comprising:a. introducing into a suitable host cell (i) an AAV vector that is free of AAV coding sequences and that comprises a heterologous gene operatively positioned between two AAV ITRs, (ii) an AAV helper construct having at least one gene encoding an AAV capsid protein, and (iii) an adenoplasmid accessory construct having a full adenoviral genome that either lacks a packaging signal or that contains sufficient additional nucleotides to be rendered unpackagable, to produce a transformed host cell;b. culturing the transformed host cell to produce replication-defective recombinant AAV virions having said heterologous gene; andc. lysing the cultured host cell to obtain said replication-defective recombinant AAV virions substantially free of wild-type AAV and adenovirus particles.