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Genetic design of an optimized packaging cell line for gene vectors transducing human B cells

Gene Therapy volume 13, pages 844–856 (2006)Cite this article

Abstract

Viral gene vectors often rely on packaging cell lines, which provide the necessary factors in trans for the formation of virus-like particles. Previously, we reported on a first-generation packaging cell line for gene vectors, which are based on the B-lymphotropic Epstein–Barr virus (EBV), a human γ-herpesvirus. This 293HEK-derived packaging cell line harbors a helper virus genome with a genetic modification that prevents the release of helper virions, but efficiently packages vector plasmids into virus-like particles with transducing capacity for human B cells. Here, we extended this basic approach towards a non-transforming, virus-free packaging cell line, which harbors an EBV helper virus genome with seven genetic alterations. In addition, we constructed a novel gene vector plasmid, which is devoid of a prokaryotic antibiotic resistance gene, and thus more suitable for in vivo applications in human gene therapy. We demonstrate in this paper that EBV-based gene vectors can be efficiently generated with this much-improved packaging cell line to provide helper virus-free gene vector stocks with transducing capacity for established human B-cell lines and primary B cells.

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Acknowledgements

We thank Joachim Ellwart and Karin Nispel for support with the cell sorting device. Our work was supported by SFB455 of the Deutsche Forschungsgemeinschaft, by the Bayerische Forschungsstiftung and by Public Health Service Grant CA70723 to WH.

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Authors and Affiliations

  1. Department of Gene Vectors, GSF-National Research Center for Environment and Health, München, Germany

    E Hettich, A Janz, D Pich, E Hellebrand & W Hammerschmidt

  2. Vaecgene Biotech GmbH, München, Germany

    E Hettich, A Janz & R Zeidler

  3. Department of Otorhinolaryngology, Clinical Cooperation Group Molecular Oncology, GSF-National Research Center for Environment and Health, München, Germany

    B Weissflog & A Moosmann

  4. Ludwig-Maximilians-Universität, München, Germany

    B Weissflog & A Moosmann

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  1. E Hettich
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  2. A Janz
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  3. R Zeidler
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  4. D Pich
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Correspondence to W Hammerschmidt.

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Hettich, E., Janz, A., Zeidler, R. et al. Genetic design of an optimized packaging cell line for gene vectors transducing human B cells. Gene Ther 13, 844–856 (2006). https://doi.org/10.1038/sj.gt.3302714

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