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Semliki Forest Virus Expression System: Production of Conditionally Infectious Recombinant Particles

Bio/Technology volume 11, pages 916–920 (1993)Cite this article

Abstract

In the recently developed Semliki Forest virus (SFV) DNA expression system, recombinant RNA encoding the viral replicase, and helper RNA molecules encoding the structural proteins needed for virus assembly are cotransfected into cells. Since the helper RNA lacks the sequence needed for its packaging into nucleocapsids, only recombinant RNAs should be packaged. We have found, however, that small amounts of replication-proficient SFV particles can still be produced. Here we describe the construction of a helper variant with a mutation in the gene encoding the viral spike protein such that its product cannot undergo normal proteolytic processing to activate viral entry functions. Hence, the recombinant stock is noninfectious, but may be activated by cleavage with chymotrypsin. When recombinant virus produced with the new helper was examined in a variety of assays, including sensitive animal tests, we were unable to detect any replication-competent SFV particles. We therefore conclude that this conditional expression system meets extremely stringent biosafety requirements.

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Author notes

  1. Peter Liljeström: Corresponding author.

Authors and Affiliations

  1. Department of Molecular Biology, Karolinska Institute, Novum, Huddinge, S-141 57, Sweden

    Peter Berglund, Mathilda Sjöberg, Henrik Garoff & Peter Liljeström

  2. Department of Microbiology, Moyne Institute, Trinity College, Dublin, 2, Ireland

    Gregory J. Atkins

  3. Department of Veterinary Pathology, Faculty of Veterinary Medicine, University College, Dublin, 4, Ireland

    Brian J. Sheahan

Authors
  1. Peter Berglund
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  2. Mathilda Sjöberg
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  4. Gregory J. Atkins
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  5. Brian J. Sheahan
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  6. Peter Liljeström
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Berglund, P., Sjöberg, M., Garoff, H. et al. Semliki Forest Virus Expression System: Production of Conditionally Infectious Recombinant Particles. Nat Biotechnol 11, 916–920 (1993). https://doi.org/10.1038/nbt0893-916

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