{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T07:34:57Z","timestamp":1768808097937,"version":"3.49.0"},"reference-count":70,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,12]],"date-time":"2021-09-12T00:00:00Z","timestamp":1631404800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Our therapeutic arsenal against viruses is very limited and the current pandemic of SARS-CoV-2 highlights the critical need for effective antivirals against emerging coronaviruses. Cellular assays allowing a precise quantification of viral replication in high-throughput experimental settings are essential to the screening of chemical libraries and the selection of best antiviral chemical structures. To develop a reporting system for SARS-CoV-2 infection, we generated cell lines expressing a firefly luciferase maintained in an inactive form by a consensus cleavage site for the viral protease 3CLPro of coronaviruses, so that the luminescent biosensor is turned on upon 3CLPro expression or SARS-CoV-2 infection. This cellular assay was used to screen a metabolism-oriented library of 492 compounds to identify metabolic vulnerabilities of coronaviruses for developing innovative therapeutic strategies. In agreement with recent reports, inhibitors of pyrimidine biosynthesis were found to prevent SARS-CoV-2 replication. Among the top hits, we also identified the NADPH oxidase (NOX) inhibitor Setanaxib. The anti-SARS-CoV-2 activity of Setanaxib was further confirmed using ACE2-expressing human pulmonary cells Beas2B as well as human primary nasal epithelial cells. Altogether, these results validate our cell-based functional assay and the interest of screening libraries of different origins to identify inhibitors of SARS-CoV-2 for drug repurposing or development.<\/jats:p>","DOI":"10.3390\/v13091814","type":"journal-article","created":{"date-parts":[[2021,9,12]],"date-time":"2021-09-12T21:48:01Z","timestamp":1631483281000},"page":"1814","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["A Bioluminescent 3CLPro Activity Assay to Monitor SARS-CoV-2 Replication and Identify Inhibitors"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6682-2029","authenticated-orcid":false,"given":"Cyrille","family":"Mathieu","sequence":"first","affiliation":[{"name":"CIRI, Centre International de Recherche en Infectiologie, Team Immunobiology of the Viral Infections, Univ Lyon, Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale (Inserm), U1111, Centre National de la Recherche Scientifique (CNRS), UMR5308, Ecole Normale Sup\u00e9rieure de Lyon, Universit\u00e9 Claude Bernard Lyon 1, 69007 Lyon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4734-2249","authenticated-orcid":false,"given":"Franck","family":"Touret","sequence":"additional","affiliation":[{"name":"Unit\u00e9 des Virus Emergents (UVE), Aix Marseille Univ, Institut de Recherche pour le D\u00e9veloppement (IRD) 190, Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale (Inserm) U1207, IHU M\u00e9diterran\u00e9e Infection, 13005 Marseille, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cl\u00e9mence","family":"Jacquemin","sequence":"additional","affiliation":[{"name":"CIRI, Centre International de Recherche en Infectiologie, Team Viral Infection, Metabolism and Immunity, Univ Lyon, Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale (Inserm), U1111, Centre National de la Recherche Scientifique (CNRS), UMR5308, Ecole Normale Sup\u00e9rieure de Lyon, Universit\u00e9 Claude Bernard Lyon 1, 69007 Lyon, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3019-9842","authenticated-orcid":false,"given":"Yves L.","family":"Janin","sequence":"additional","affiliation":[{"name":"Unit\u00e9 de Chimie et Biocatalyse, Institut Pasteur, Centre National de la Recherche Scientifique (CNRS), UMR 3523, 28 rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antoine","family":"Nougair\u00e8de","sequence":"additional","affiliation":[{"name":"Unit\u00e9 des Virus Emergents (UVE), Aix Marseille Univ, Institut de Recherche pour le D\u00e9veloppement (IRD) 190, Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale (Inserm) U1207, IHU M\u00e9diterran\u00e9e Infection, 13005 Marseille, 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