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Transfection-independent production of alphavirus replicon particles based on poxvirus expression vectors

Nature Biotechnology volume 21, pages 932–935 (2003)Cite this article

Abstract

This report describes a transfection-independent system for packaging alphavirus replicon vectors using modified vaccinia virus Ankara (MVA) vectors to express all of the RNA components necessary for the production of Venezuelan equine encephalitis (VEE) virus replicon particles (VRP). Infection of mammalian cells with these recombinant MVA vectors resulted in robust expression of VEE structural genes, replication of the alphavirus vector and high titers of VRP. In addition, VRP packaging was achieved in a cell type (fetal rhesus lung) that has been approved for the manufacturing of vaccines destined for human use.

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Figure 1: MVA-based VRP packaging system.

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Acknowledgements

We thank Bernard Moss and Linda Wyatt for providing the vaccinia virus vector plasmids and MVA, and Jonathan Smith, Mike Parker, Peter Pushko and Kurt Kamrud for helpful discussions and technical advice with the VEE expression system.

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Author notes

  1. Nikos Vasilakis

    Present address: Department of Pathology, University of Texas Medical Branch at Galveston, Galveston, Texas, 77555, USA

  2. Gerald R Kovacs

    Present address: Office of Biodefense Research Affairs, National Institute of Allergy and Infectious Diseases, National Institutes of Heath, Bethesda, Maryland, 20892, USA

Authors and Affiliations

  1. Viral Vaccine Discovery, Wyeth Vaccines Research, Pearl River, 10965, New York, USA

    Nikos Vasilakis, Darlene Falvey, Seema S Gangolli, John Coleman, Jacek Kowalski, Stephen A Udem, Timothy J Zamb & Gerald R Kovacs

  2. Department of Pathology, University of Texas Medical Branch at Galveston, Galveston, Texas, 77555, USA

    Gerald R Kovacs

  3. Office of Biodefense Research Affairs, National Institute of Allergy and Infectious Diseases, National Institutes of Heath, Bethesda, Maryland, 20892, USA

    Nikos Vasilakis

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  1. Nikos Vasilakis
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Correspondence to Gerald R Kovacs.

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Vasilakis, N., Falvey, D., Gangolli, S. et al. Transfection-independent production of alphavirus replicon particles based on poxvirus expression vectors. Nat Biotechnol 21, 932–935 (2003). https://doi.org/10.1038/nbt845

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