Use of adeno-associated virus as a mammalian DNA cloning vector: transduction of neomycin resistance into mammalian tissue culture cells

Abstract

Adeno-associated virus (AAV) is a human DNA virus that has a broad host range and can be grown both as an integrated provirus and as a lytic genome. These properties suggested that AAV may be useful as a mammalian transduction vector. To test this possibility, we have isolated a recombinant AAV viral stock in which the neomycin resistance gene was substituted for the AAV capsid genes. Using this recombinant stock, we have demonstrated that AAV can be used to transduce foreign DNA into human and murine tissue culture cells. In addition, we have demonstrated that, if the transductants are superinfected with a helper virus (adenovirus), the recombinant AAV genome is rescued from the proviral state and amplified to high copy number. These unique features of AAV vectors suggest that they may have a broad utility in the study of biological problems. Because AAV, itself, is nonpathogenic in both humans and animals, these vectors also may be useful for the purpose of gene therapy.