Email this article Generation and characterization of SARS-CoV-2 pseudoviruses
- Authors: Rudometova N.B.1, Shcherbakov D.N.1, Karpenko L.I.1
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Affiliations:
- State Research Center of Virology and Biotechnology “Vector”, Federal Service for Surveillance on Consumer Rights Protection and Human Well-being
- Issue: Vol 22, No 2 (2022)
- Pages: 249-253
- Section: Conference proceedings
- URL: https://journals.rcsi.science/MAJ/article/view/108600
- DOI: https://doi.org/10.17816/MAJ108600
- ID: 108600
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Abstract
BACKGROUND: Pseudovirus technology is a versatile and valuable tool for both fundamental and applied virological research. Pseudotyped viruses provide the same cell entry mechanism as SARS-CoV-2 and are widely used to investigate the virus entry mechanism, cell tropism, and virus neutralization assays.
AIM: The aim of the work is to obtain pseudotyped SARS-CoV-2 viruses and evaluate their transducing activity.
MATERIALS AND METHODS: Using genetic engineering methods, a genetic construct carrying the SARS-CoV-2 glycoprotein S gene was obtained, as well as the pLenti-Luc-GFP reporter plasmid encoding the green fluorescent protein (GFP) and firefly luciferase genes. Pseudovirus particles were generated by transfection of eukaryotic cells. The transducing activity of pseudoviral particles displaying SARS-CoV-2 glycoprotein S on their surface was studied using HEK293, HEK293-hACE2, and HEK293-hACE2-TMPRSS2 (t) cell cultures.
RESULTS: Based on the second-generation lentiviral platform, pseudoviruses were obtained that exhibit SARS-CoV-2 S glycoprotein on their surface. It was found that the pseudoviruses penetrate more efficiently into HEK293-hACE2-TMPRSS2 cells than into HEK293-hACE2. Pseudoviruses have been shown to be sensitive to neutralization by recombinant monoclonal antibodies that interact with the receptor-binding domain (RBD) of the SARS-CoV-2 S glycoprotein.
CONCLUSIONS: The pseudoviruses can be used both to search for antiviral drugs that would be able to block the penetration of SARS-CoV-2 into the target cell, and to evaluate the effectiveness of the developed monoclonal antibodies and vaccines against SARS-CoV-2.
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##article.viewOnOriginalSite##About the authors
Nadezhda B. Rudometova
State Research Center of Virology and Biotechnology “Vector”, Federal Service for Surveillance on Consumer Rights Protection and Human Well-being
Author for correspondence.
Email: nadenkaand100@mail.ru
ORCID iD: 0000-0002-1684-9071
SPIN-code: 5283-6608
Cand. Sci. (Biol.), Research Associate
Russian Federation, Koltsovo, Novosibirsk RegionDmitry N. Shcherbakov
State Research Center of Virology and Biotechnology “Vector”, Federal Service for Surveillance on Consumer Rights Protection and Human Well-being
Email: dnshcherbakov@gmail.com
ORCID iD: 0000-0001-8023-4453
SPIN-code: 9616-0933
Scopus Author ID: 37027118800
Dr. Sci. (Biol.), Leading Research Associate
Russian Federation, Koltsovo, Novosibirsk RegionLarisa I. Karpenko
State Research Center of Virology and Biotechnology “Vector”, Federal Service for Surveillance on Consumer Rights Protection and Human Well-being
Email: karpenko@vector.nsc.ru
ORCID iD: 0000-0003-4365-8809
SPIN-code: 2026-5992
Scopus Author ID: 7005000410
Dr. Sci. (Biol.), Assistant Professor, Leading Research Associate
Russian Federation, Koltsovo, Novosibirsk RegionReferences
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