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Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
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Reverse Genetics System for Rabbit vesivirus.

Ángel L Álvarez1, Alberto García-Manso1, Kevin P Dalton1

  • 1Departamento de Bioquímica y Biología Molecular, Instituto Universitario de Biotecnología de Asturias (IUBA), Universidad de Oviedo, Oviedo, Spain.

Frontiers in Microbiology
|December 11, 2020
PubMed
Summary

Researchers developed a novel reverse genetics system for Rabbit vesivirus (RaV), a calicivirus. This system enables efficient virus rescue from cDNA, facilitating studies on calicivirus replication and pathogenesis.

Keywords:
Vesiviruscalicivirusinfectious cDNA clonereverse geneticsvirus rescue

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Area of Science:

  • Virology
  • Molecular Biology

Background:

  • Most caliciviruses are difficult to grow in cell culture.
  • Rabbit vesivirus (RaV) is an exception, replicating efficiently in various cell lines.
  • Efficient replication makes RaV suitable for reverse genetics studies.

Purpose of the Study:

  • To establish an efficient and consistent RaV rescue system using a cDNA transcription vector.
  • To enable reverse genetics studies for understanding calicivirus biology.

Main Methods:

  • Construction of a RaV infectious cDNA clone using a plasmid expression vector.
  • Transfection of permissive cells with the plasmid DNA and T7 RNA-polymerase.
  • In vitro transcription of genome-length RNAs with 5'-cap addition, followed by transfection.

Main Results:

  • Successful de novo synthesis of viral RNA transcripts and recovery of infectious RaV progeny.
  • Demonstration of typical virus-induced cytopathic effects.
  • Efficient virus recovery using both in trans and in vitro transcription methods.

Conclusions:

  • An efficient RaV rescue system based on a cDNA transcription vector has been established.
  • This system serves as a valuable tool for investigating calicivirus biology through reverse genetics.
  • The findings pave the way for deeper understanding of calicivirus replication and pathogenesis.