A protocol for displaying viral envelope glycoproteins on the surface of vesicular stomatitis viruses

STAR Protoc. 2020 Dec 9;1(3):100209. doi: 10.1016/j.xpro.2020.100209. eCollection 2020 Dec 18.

Abstract

We describe the production of single-cycle (sc) and replication-competent recombinant vesicular stomatitis viruses (rcVSVs) displaying heterologous envelope glycoproteins (Envs) on their surface. We prepare scVSVs by transiently expressing HIV-1 Envs or SARS-CoV-2 spike followed by infection of the cells with scVSV particles, which do not carry the vsv-g gene. To prepare rcVSVs, we replace the vsv-g with a specific env-encoding gene, transfect cells with multiple plasmids for production of the genomic RNA and viral proteins, and rescue replication-competent viruses.

Keywords: Cell culture; Cell-based assays; Immunology; Molecular biology.

Publication types

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

MeSH terms

  • Animals
  • COVID-19 / virology
  • Cell Line
  • Cricetinae
  • HIV-1 / genetics
  • Humans
  • Protein Engineering
  • Recombinant Proteins* / chemistry
  • Recombinant Proteins* / genetics
  • Recombinant Proteins* / metabolism
  • SARS-CoV-2 / genetics
  • Spike Glycoprotein, Coronavirus* / chemistry
  • Spike Glycoprotein, Coronavirus* / genetics
  • Spike Glycoprotein, Coronavirus* / metabolism
  • Vesicular Stomatitis / genetics*
  • env Gene Products, Human Immunodeficiency Virus* / chemistry
  • env Gene Products, Human Immunodeficiency Virus* / genetics
  • env Gene Products, Human Immunodeficiency Virus* / metabolism

Substances

  • Recombinant Proteins
  • Spike Glycoprotein, Coronavirus
  • env Gene Products, Human Immunodeficiency Virus
  • spike protein, SARS-CoV-2