A method for producing helper-free stocks of recombinant adeno-associated virus
(rAAV) which can be used to efficiently and stably transduce foreign genes into
host cells or organisms. The method comprises the cotransfection of eukaryotic
cells with rAAV and with helper AAV DNA in the presence of helper virus (e.g. adenovirus
or herpesvirus) such that the helper AAV DNA is not associated with virion formation.
The crux of the invention lies in the inability of the helper AAV DNA to recombine
with rAAV vector, thereby preventing the generation of wild-type virus. The pure
stocks of recombinant AAV provide an AAV viral expression vector system with increased
yield of recombinant virus, improved efficiency, higher definition, and greater
safety than presently used systems.