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J Gen Virol 85 (2004), 2209-2214; DOI 10.1099/vir.0.79940-0

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© 2004 Society for General Microbiology

Short Communication

The adenovirus E1A and E1B19K genes provide a helper function for transfection-based adeno-associated virus vector production

Takashi Matsushita1, Takashi Okada1, Toshiya Inaba2, Hiroaki Mizukami1, Keiya Ozawa1 and Peter Colosi3

1 Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, 3311-1 Yakushiji, Minami-kawachi, Kawachi, Tochigi 329-0489, Japan
2 Department of Molecular Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan
3 Avigen Inc., Alameda, CA, USA

Correspondence
Takashi Okada
tokada{at}jichi.ac.jp
Peter Colosi
PColosi{at}avigen.com

Although the adenoviral E1, E2A, E4 and VA RNA regions are required for efficient adeno-associated virus (AAV) vector production, the role that the individual E1 genes (E1A, E1B19K, E1B55K and protein IX) play in AAV vector production has not been clearly determined. E1 mutants were analysed for their ability to mediate AAV vector production in HeLa or KB cells, when cotransfected with plasmids encoding all other packaging functions. Disruption of E1A and E1B19K genes resulted in vector yield reduction by up to 10- and 100-fold, respectively, relative to the wild-type E1. Interruption of the E1B55K and protein IX genes had a modest effect on vector production. Interestingly, expression of anti-apoptotic E1B19K cellular homologues such as Bcl-2 or Bcl-xL fully complemented E1B19K mutants for AAV vector production. These findings may be valuable for the future development of packaging cell lines for AAV vector production.






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