Production of recombinant adeno-associated virus vectors

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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4353201, 935 27, 935 32, 935 56, 935 57, C12N 1564, C12N 1585, C12N 1586, C12N 1565

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active

051734141

ABSTRACT:
A simplified method to produce recombinant adeno-associated virus (AAV) vectors is described. The procedure involves the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr Virus (oriP), and a recombinant AAV genome. The chimeric plasmids themselves are also a part of the present invention. These EBV/AAV plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of recombinant AAV virus upon addition of wild-type, adeno-associated virus helper functions. Recombinant AAV vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit all of the attributes of viral elements produced by conventional methods.

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