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.
© 2020 The Author(s).
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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COVID-19 / virology
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Cell Line
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Cricetinae
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HIV-1 / genetics
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Humans
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Protein Engineering
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Recombinant Proteins* / chemistry
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Recombinant Proteins* / genetics
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Recombinant Proteins* / metabolism
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SARS-CoV-2 / genetics
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Spike Glycoprotein, Coronavirus* / chemistry
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Spike Glycoprotein, Coronavirus* / genetics
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Spike Glycoprotein, Coronavirus* / metabolism
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Vesicular Stomatitis / genetics*
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env Gene Products, Human Immunodeficiency Virus* / chemistry
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env Gene Products, Human Immunodeficiency Virus* / genetics
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env Gene Products, Human Immunodeficiency Virus* / metabolism
Substances
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Recombinant Proteins
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Spike Glycoprotein, Coronavirus
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env Gene Products, Human Immunodeficiency Virus
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spike protein, SARS-CoV-2