Replication and single-cycle delivery of SARS-CoV-2 replicons

Science. 2021 Nov 26;374(6571):1099-1106. doi: 10.1126/science.abj8430. Epub 2021 Oct 14.

Abstract

Molecular virology tools are critical for basic studies of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and for developing new therapeutics. Experimental systems that do not rely on viruses capable of spread are needed for potential use in lower-containment settings. In this work, we use a yeast-based reverse genetics system to develop spike-deleted SARS-CoV-2 self-replicating RNAs. These noninfectious self-replicating RNAs, or replicons, can be trans-complemented with viral glycoproteins to generate replicon delivery particles for single-cycle delivery into a range of cell types. This SARS-CoV-2 replicon system represents a convenient and versatile platform for antiviral drug screening, neutralization assays, host factor validation, and viral variant characterization.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral / immunology
  • Antiviral Agents / pharmacology
  • Cell Line
  • Humans
  • Interferons / pharmacology
  • Microbial Sensitivity Tests
  • Mutation
  • Plasmids
  • RNA, Viral / genetics*
  • RNA, Viral / metabolism
  • Replicon / genetics
  • Replicon / physiology*
  • Reverse Genetics
  • SARS-CoV-2 / drug effects
  • SARS-CoV-2 / genetics*
  • SARS-CoV-2 / physiology
  • Saccharomyces cerevisiae / genetics
  • Spike Glycoprotein, Coronavirus / genetics
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism
  • Viral Pseudotyping
  • Virion / genetics
  • Virion / physiology
  • Virus Replication

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Antiviral Agents
  • NSP1 protein, SARS-CoV-2
  • RNA, Viral
  • Spike Glycoprotein, Coronavirus
  • Viral Nonstructural Proteins
  • spike protein, SARS-CoV-2
  • Interferons