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Related Experiment Videos

Recombinant vesicular stomatitis viruses from DNA

N D Lawson1, E A Stillman, M A Whitt

  • 1Department of Pathology, Yale University School of Medicine, New Haven, CT 06510, USA.

Proceedings of the National Academy of Sciences of the United States of America
|May 9, 1995
PubMed
Summary

Scientists created a vesicular stomatitis virus (VSV) from a DNA clone, enabling genetic engineering for potential vaccines. This breakthrough allows for easier VSV replication studies and development of novel viral therapies.

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Vesicular stomatitis virus (VSV) is a prototype rhabdovirus with a positive-strand RNA genome.
  • Genetic manipulation of VSV has been challenging due to its RNA genome.

Purpose of the Study:

  • To develop a method for generating infectious VSV from a DNA clone.
  • To enable genetic analysis and engineering of VSV for potential applications.

Main Methods:

  • Assembly of a DNA clone encoding the full-length VSV genome.
  • In vitro transcription using bacteriophage T7 RNA polymerase.
  • Expression of viral RNA and proteins in cultured cells for virus recovery.

Main Results:

  • Successfully recovered infectious VSV with wild-type growth characteristics from the DNA clone.

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  • Verified virus recovery through genetic tagging, RNA sequencing, and recombinant virus production.
  • Demonstrated the feasibility of generating VSV recombinants with modified glycoproteins.

    • Conclusions:

    • The ability to generate VSV from DNA facilitates genetic analysis of viral replication.
    • Genetically engineered VSV displaying foreign antigens can be produced, offering potential as vaccines against other viruses.