Lentivirus production is influenced by SV40 large T-antigen and chromosomal integration of the vector in HEK293 cells.
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Authors
Gama-Norton, LeonorBotezatu, Lacramioara
Herrmann, Sabrina
Schweizer, Matthias
Alves, Paula Marques
Hauser, Hansjoerg
Wirth, Dagmar
Issue Date
2011-10
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Show full item recordAbstract
Currently, lentiviral vectors for research and gene therapy are produced from 293-T cells that are transiently transfected with plasmids encoding the vector and helper functions. However, transiently transfected vectors as well as the presence of SV40 virus large T-antigen (T-Ag) cause serious technical and safety considerations. We aimed to exploit single copy integration sites in the HEK293 genome supporting lentiviral vector production. We found that lentiviral vectors result in minimal infectious particle production from single copy integrants in HEK293. Moreover, once this cell line harbors single copy integrations of lentiviral vectors, its ability to transiently produce lentiviral vectors becomes strongly impaired. T-Ag has a dramatic effect on virus production. Low levels of constitutive T-Ag expression can overcome the production restriction imposed by integrated lentiviral vectors copies. Interestingly, T-Ag does not exert its role at the level of transcriptional activity of the vector; rather, it seems to impose an indirect effect on the cell thereby enabling lentiviral vector production. Altogether, our study highlights the restrictions for integrated lentiviral vectors that are relevant for the establishment of stable and safe producer cell lines.Citation
Lentivirus production is influenced by SV40 large T-antigen and chromosomal integration of the vector in HEK293 cells. 2011, 22 (10):1269-79 Hum. Gene Ther.Affiliation
Helmholtz Centre for Infection Research, Braunschweig, Germany .Journal
Human gene therapyPubMed ID
21554103Type
ArticleLanguage
enISSN
1557-7422ae974a485f413a2113503eed53cd6c53
10.1089/hum.2010.143
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