Generation of recombinant adeno-associated virus vectors by a complete adenovirus-mediated approach.
An efficient approach to the large-scale production of rAAV will significantly facilitate the application of this promising gene delivery vector in human gene therapy applications. In this study, we report a novel approach to rAAV production that is based exclusively on the adenovirus vector, without the need for plasmid transfections and special packaging cell lines. All components required for rAAV production, including the rep and cap genes, and the therapeutic gene(s) are delivered to the widely used 293 packaging cell line by adenovirus vectors. High-titer rAAV vectors (200-600 infectious units/producer cell) were obtained by use of this approach. As adenovirus vectors can be produced to high titers and they can infect cells in suspension efficiently, this approach may be amenable to scaled-up production of rAAV vectors.
Zhang
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Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710, USA.
Li
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D000256:Adenoviridae / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D014162:Transfection
D000900:Anti-Bacterial Agents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004318:Doxycycline