The purpose of this work was to develop an efficient Therefore the minimum set of genes required to produce method for the production of adeno-associated virus (AAV) AAV helper activity equivalent to that provided by adenvectors in the absence of helper virus. The adenovirus ovirus infection consists of, or is a subset of, the following regions that mediate AAV vector replication were identified genes: the E4orf6 gene, the 72-M r , E2A protein gene, the and assembled into a helper plasmid. These included the VA RNA genes and the E1 region. AAV vector preparations VA, E2A and E4 regions. When this helper plasmid was made with adenovirus and by the helper virus-free method cotransfected into 293 cells, along with plasmids encoding were essentially indistinguishable with respect to particle the AAV vector, and rep and cap genes, AAV vector was density, particle to infectivity ratio, capsimer ratio and produced as efficiently as when using adenovirus infection efficiency of muscle transduction in vivo. Only AAV vector as a source of help. CMV-driven constructs expressing the preparations made by the helper virus-free method were E4orf6 and the 72-M r , E2A proteins were able to funcnot reactive with anti-adenovirus sera. tionally replace the E4 and E2A regions, respectively.

Mucopolysaccharidosis type VII (MPS VII) is a lysosomalproduced low GUS activity in several tissues. This latter storage disease caused by a genetic deficiency of ␤-glucutreatment of MPS VII mice reduced glycosaminoglycan ronidase (GUS). We used a recombinant adeno-associalevels in the liver to normal and reduced storage granules ted virus vector (AAV-GUS) to deliver GUS cDNA to MPS dramatically. We show that a single administration of VII mice. The route of vector administration had a dramatic AAV-GUS can provide sustained expression of GUS in a effect on the extent and distribution of GUS activity. Intravariety of cell types and is sufficient to reverse the disease muscular injection of AAV-GUS resulted in high, localized phenotype at least in the liver. production of GUS, while intravenous administration