A novel method using baculovirus-mediated gene transfer for production of recombinant adeno-associated virus vectors.
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.
Sollerbrant
Kerstin
K
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.|Center for Genomics Research, Karolinska Institute, SE-17177 Stockholm, Sweden1.
Elmén
Joacim
J
Center for Genomics Research, Karolinska Institute, SE-17177 Stockholm, Sweden1.
Wahlestedt
Claes
C
Center for Genomics Research, Karolinska Institute, SE-17177 Stockholm, Sweden1.
Acker
Joel
J
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.
Leblois-Prehaud
Hélene
H
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.
Latta-Mahieu
Martine
M
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.
Yeh
Patrice
P
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.
Perricaudet
Michel
M
CNRS UMR 1582, Vectorologie et Transfert de Génes, Institut Gustave Roussy, 94805 Villejuif Cedex, France2.
D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D011995:Recombination, Genetic; D014779:Virus Replication