Production of Recombinant Adeno‐Associated Viral Vectors

Recombinant adeno‐associated viral (rAAV) vectors are capable of delivering genes to both dividing and nondividing cells. This property, along with the physical stability of the virions, permits transduction of cells both in tissue culture and in the whole animal. In this unit, protocols for the adenovirus‐free production of rAAV vectors, their purification, and their quantitation are provided. The production and purification of the rAAV vectors are described, using two different methods. Two assays for determining rAAV titers are also presented: dot blot and in vitro infection with a transgene expression assay.