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.
Original language | English (US) |
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Article number | 100209 |
Journal | STAR Protocols |
Volume | 1 |
Issue number | 3 |
DOIs | |
State | Published - Dec 18 2020 |
Bibliographical note
Funding Information:We thank Jason McLellan (University of Texas at Austin) for providing the plasmid for the expression of soluble SARS-CoV-2 spike, Stefan P?hlmann (University G?ttingen) for providing the plasmid for the expression of SARS-CoV-2 spike, Ryan Langlois (University of Minnesota) for the expression plasmids of VSV proteins, Fang Li (University of Minnesota) for providing the 293T-ACE2 cells, and Stefan P?hlmann (University G?ttingen ). This work was supported by internal funds of the Department of Medicine at the University of Minnesota and by NIH/NIDA grant 1DP2DA049279-01 (to A.H.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. T.A.F. and A.H. wrote the protocol, A.C. and T.A.F. prepared the figures, and all authors reviewed the manuscript. The authors declare no competing interests.
Funding Information:
We thank Jason McLellan (University of Texas at Austin) for providing the plasmid for the expression of soluble SARS-CoV-2 spike, Stefan Pӧhlmann (University Gӧttingen) for providing the plasmid for the expression of SARS-CoV-2 spike, Ryan Langlois (University of Minnesota) for the expression plasmids of VSV proteins, Fang Li (University of Minnesota) for providing the 293T-ACE2 cells, and Stefan Pӧhlmann (University Gӧttingen ). This work was supported by internal funds of the Department of Medicine at the University of Minnesota and by NIH/ NIDA grant 1DP2DA049279-01 (to A.H.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Publisher Copyright:
© 2020 The Author(s)
Keywords
- Cell culture
- Cell-based assays
- Immunology
- Molecular biology
PubMed: MeSH publication types
- Journal Article