{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T09:45:55Z","timestamp":1753868755537,"version":"3.41.2"},"reference-count":14,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2024,11,21]],"date-time":"2024-11-21T00:00:00Z","timestamp":1732147200000},"content-version":"vor","delay-in-days":20,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2024,11]]},"abstract":"Abstract<\/jats:title>Protease inhibitors are among the most powerful antiviral drugs. They have been used successfully against viruses, such as the human immunodeficiency virus (HIV), hepatitis C virus (HCV) and severe acute respiratory syndrome coronavirus 2 (SARS\u2010CoV\u20102). Protease inhibitor screening tools are therefore important to identify inhibitors that have the potential to become antiviral drugs. In this article, we describe newly developed cell\u2010 and virus replicon\u2010based platforms to screen inhibitors. We developed the methods presented here by genetically modifying vesicular stomatitis virus, a model virus from the family Rhabdoviridae<\/jats:italic>. \u00a9 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.<\/jats:p>Basic Protocol<\/jats:bold>: Mpro<\/jats:sup>\u2010On and \u2010Off assay<\/jats:p>Alternate Protocol 1<\/jats:bold>: Virus production with transient P\u2010 and L TransIT transfection<\/jats:p>Alternate Protocol 2<\/jats:bold>: Virus production with transient P\u2010 and L Ca2<\/jats:sub>PO4<\/jats:sub> transfection<\/jats:p>Alternate Protocol 3<\/jats:bold>: Luciferase\u2010based variation of the On assay<\/jats:p>Alternate Protocol 4<\/jats:bold>: Screening assay with fluorescence\u2010activated cell sorting readout<\/jats:p>Support Protocol<\/jats:bold>: Performing kinetic measurements with Off assay<\/jats:p>","DOI":"10.1002\/cpz1.70062","type":"journal-article","created":{"date-parts":[[2024,11,21]],"date-time":"2024-11-21T15:29:27Z","timestamp":1732202967000},"source":"Crossref","is-referenced-by-count":0,"title":["Vesicular Stomatitis Virus as a Platform for Protease Activity Measurements"],"prefix":"10.1002","volume":"4","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-7415-0211","authenticated-orcid":false,"given":"Stefanie","family":"Rauch","sequence":"first","affiliation":[{"name":"Institute of Virology Medical University of Innsbruck Innsbruck Austria"},{"name":"These authors contributed equally to this work"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco","family":"Costacurta","sequence":"additional","affiliation":[{"name":"Institute of Virology Medical University of Innsbruck Innsbruck Austria"},{"name":"These authors contributed equally to this work"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dorothee","family":"von Laer","sequence":"additional","affiliation":[{"name":"Institute of Virology Medical University of Innsbruck Innsbruck Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8148-9490","authenticated-orcid":false,"given":"Emmanuel","family":"Heilmann","sequence":"additional","affiliation":[{"name":"Institute of Virology Medical University of Innsbruck Innsbruck Austria"},{"name":"Biological and Environmental Science and Engineering Division King Abdullah University of Science and Technology Thuwal Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,11,21]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.73.5.1504"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.73.2.442"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.3824\/stembook.1.85.1"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2000.0736"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00211\u201006"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s42003\u2010022\u201003277\u20100"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.3390\/v11110989"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010021\u201021630\u20105"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01371\u201015"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.15.4.1004\u20101011.1975"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00109\u2010012\u20100863\u20106"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1006\/smvy.1997.0105"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.4.2318\u20102323.1996"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.78.11.5670\u20105678.2004"}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.70062","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T04:58:06Z","timestamp":1733115486000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.70062"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11]]},"references-count":14,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2024,11]]}},"alternative-id":["10.1002\/cpz1.70062"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.70062","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"type":"print","value":"2691-1299"},{"type":"electronic","value":"2691-1299"}],"subject":[],"published":{"date-parts":[[2024,11]]},"article-number":"e70062"}}