Cell lines for the production of recombinant adeno-associated virus

Human Gene Therapy
K R ClarkP R Johnson

Abstract

Adeno-associated virus (AAV) is a replication-defective parvovirus that is being developed as a vector for human gene transfer. However, a major obstacle to commonplace usage of AAV vectors is the production of recombinant virions (rAAV) in sufficient quantities for not only human trials, but also for preclinical studies of basic biology, toxicology, and efficacy. Unfortunately, current methods for large-scale production are cumbersome and expensive. We have developed a simplified method for generating rAAV by establishing neomycin-resistant cell lines containing copies of the AAV rep-cap genes and a rAAV vector. After infection with adenovirus, these cell lines are shown to produce infectious rAAV in relatively high titer. This method eliminates the need for exogenous DNA transfection and scale-up procedures are limited only by the normal constraints of growing cells in culture.

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Apr 14, 2006·Human Genetics·Kenneth H Warrington, Roland W Herzog
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