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

• Published in: STAR protocols Vol. 1; no. 3; p. 100209
• Main Authors: Farzani, Touraj Aligholipour, Chov, Angela, Herschhorn, Alon
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.12.2020; Elsevier
• Subjects: Animals; Cell culture; Cell Line; Cell-based assays; COVID-19 - virology; Cricetinae; env Gene Products, Human Immunodeficiency Virus - chemistry; env Gene Products, Human Immunodeficiency Virus - genetics; env Gene Products, Human Immunodeficiency Virus - metabolism; HIV-1 - genetics; Humans; Immunology; Molecular biology; Protein Engineering; Protocol; 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; Cell culture; Immunology; Molecular biology; Cell-based assays
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2020.100209

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. [Display omitted] •Heterologous, viral envelope glycoproteins can be displayed on recombinant VSV•Recombinant VSVs allow study of the biology of viral entry of different viruses•Recombinant VSVs can be used to measure virus neutralization and as vaccines 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.