1] Adeno-associated virus: Integrating vectors for human gene therapy

Abstract

Publisher Summary This chapter describes the generation of a recombinant adeno-associated virus (rAAV) that expresses the Escherichia coli β-galactosidase ( lac Z) gene and its use in identifying cell types permissive to AAV infection. Viral research has revealed a number of viruses that can be modified to introduce novel genes efficiently into a variety of cell types, including adenovirus, retrovirus, vaccinia virus, herpesvirus, and adeno-associated virus (AAV). Some of the unique features that make this virus attractive for gene therapy include the facts that AAV is prevalent in humans, it has never been identified as a causative agent of human disease, and it is able to insert its genome locus-specifically into human chromosomes. AAVs are members of the family Parvoviridiae in the genus dependovirus , appropriately named for their complete dependence on coinfection with a helper virus for productive infection. In the absence of helper virus, the wild-type ( wt ) AAV genome integrates efficiently in a locus-specific manner into the cellular genome and can exist as a provirus for many cellular generations until rescue with a helper virus. AAVs are among the smallest DNA animal viruses. The production of recombinant lacZ AAV involves cotransfection of the AAV vector and helper plasmids into permissive cells, followed by adenovirus infection.