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A reporter cell line for the automated quantification of SARS-CoV-2 infection in living cells.

Authors :
Desmarets L
Callens N
Hoffmann E
Danneels A
Lavie M
Couturier C
Dubuisson J
Belouzard S
Rouillé Y
Source :
Frontiers in microbiology [Front Microbiol] 2022 Sep 29; Vol. 13, pp. 1031204. Date of Electronic Publication: 2022 Sep 29 (Print Publication: 2022).
Publication Year :
2022

Abstract

The SARS-CoV-2 pandemic and the urgent need for massive antiviral testing highlighted the lack of a good cell-based assay that allowed for a fast, automated screening of antivirals in high-throughput content with minimal handling requirements in a BSL-3 environment. The present paper describes the construction of a green fluorescent substrate that, upon cleavage by the SARS-CoV-2 main protease, re-localizes from the cytoplasm in non-infected cells to the nucleus in infected cells. The construction was stably expressed, together with a red fluorescent nuclear marker, in a highly susceptible clone derived from Vero-81 cells. With this fluorescent reporter cell line, named F1G-red, SARS-CoV-2 infection can be scored automatically in living cells by comparing the patterns of green and red fluorescence signals acquired by automated confocal microscopy in a 384-well plate format. We show the F1G-red system is sensitive to several SARS-CoV-2 variants of concern and that it can be used to assess antiviral activities of compounds in dose-response experiments. This high-throughput system will provide a reliable tool for antiviral screening against SARS-CoV-2.<br />Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2022 Desmarets, Callens, Hoffmann, Danneels, Lavie, Couturier, Dubuisson, Belouzard and Rouillé.)

Details

Language :
English
ISSN :
1664-302X
Volume :
13
Database :
MEDLINE
Journal :
Frontiers in microbiology
Publication Type :
Academic Journal
Accession number :
36246297
Full Text :
https://doi.org/10.3389/fmicb.2022.1031204