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J Gen Virol. 2001 Sep;82(Pt 9):2051-60.

A novel method using baculovirus-mediated gene transfer for production of recombinant adeno-associated virus vectors.

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  • 1Center for Genomics Research, Karolinska Institute, SE-17177 Stockholm, Sweden. kerstin.sollerbrant@licr.ki.se

Abstract

The baculovirus Autographa californica multiple nucleopolyhedrosis virus causes non-productive infection in mammalian cells. Recombinant baculovirus therefore has the capability to transfer and express heterologous genes in these cells if a mammalian promoter governs the gene of interest. We have investigated the possibility of using baculovirus as a tool to produce recombinant adeno-associated virus (rAAV). AAV has become increasingly popular as a vector for gene therapy and functional genomics efforts, although its use is hampered by the lack of a simple and efficient vector production method. We show here that co-infection of mammalian producer cells with three viruses - a baculovirus containing the reporter gene flanked by AAV ITRs, a baculovirus expressing the AAV rep gene and a helper adenovirus expressing the AAV cap gene - produces infectious rAAV particles. This baculovirus-based chimeric vector method may in future improve large-scale rAAV vector preparations and circumvent present-day problems associated with rAAV production.

PMID:
11514714
[PubMed - indexed for MEDLINE]
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