In vitro packaging of an infectious recombinant adeno-associated virus 2

Adeno-associated virus 2 (AAV), a human parvovirus, has properties such as stable chromosomal integration, high infectivity and lack of known human pathogenicity, making it a potentially useful vector for human gene therapy. AAV requires a helper virus, such as an adenovirus, for optimal replication and packaging in mammalian cells. Although replication of the wild-type AAV genome has been demonstrated in vitro, packaging of infectious viral particles was not documented until now. In this study, we produced in vitro infectious recombinant AAV virions containing the neomycin resistance (NeoR) and the human CD-16 (FCγRIIIa) gene. Our in vitro packaging of AAV has the same characteristics as AAV produced in vivo and demonstrates the feasibility of developing a safe packaging method for AAV to be used in gene therapy.