Abstract: Disclosed are HSV-1 amplicons that supply all necessary helper functions required for rAAV packaging and methods for their use. These HSV-1 amplicons have been shown to be capable of rescuing and replicating all forms of rAAV genomes including rAAV genomes introduced into cells by infection of rAAV virions, rAAV genomes transfected into cells on plasmids or proviral rAAV genomes integrated into cellular chromosomal DNA. Also provided are methods for preparing high-titer rAAV vector compositions suitable for gene therapy and the delivery of exogenous polynucleotides to selected host cells.

Abstract: A human gamma herpesviris genome known as Kaposi sarcoma associated herpesvirus (KSHV) or human herpesvirus 8 (HHV-8) is present in virtually all AIDS and non-AIDS Kaposi's sarcoma (KS) lesions, as well as in body cavity based lymphomas (BCBL), Multiple myeloma, and in multicentric Casdeman's disease. Isolation and DNA sequencing of a 17-kb segment encompassing a HHV-8 divergent locus (DL-B) between ORF11 and ORF17 revealed the presence of nine viral ORFs with gene products related to cellular proteins. These include the complete thymidylate synthase (TS) gene and a dihydrofolate reductase (DHFR) gene, four cytokine genes (vIL6, vMIP-1A, vMIP-1B and BCK) that have not previously been found to be encoded by a virus, and a Bcl2 homologue. This region in HHV-8 also contains the T1.1 abundant lytic cycle nuclear RNA gene and encompasses two genes (or exons) encoding proteins with C4HC3 zinc finger domains of the PHD/LAP subtype.