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There is a need for an efficient inhibitor screening system that can identify drug candidates in a cellular context. Here we describe modular, tunable autoproteolytic gene switches (TAGS) relying on synthetic transcription factors that self-inactivate,\u00a0unless in the presence of coronavirus protease inhibitors, consequently activating transgene expression. TAGS rapidly report the impact of drug candidates on Mpro and PLpro activities with a high signal-to-noise response and a sensitivity matching concentration ranges inhibiting viral replication. The modularity of the TAGS enabled the study of other <jats:italic>Coronaviridae<\/jats:italic> proteases, characterization of mutations and multiplexing of gene switches in human cells. Mice implanted with Mpro or PLpro TAGS-engineered cells enabled analysis of the activity and bioavailability of protease inhibitors in vivo in a virus-free setting.<\/jats:p>","DOI":"10.1038\/s41467-021-27072-3","type":"journal-article","created":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T11:02:58Z","timestamp":1637578978000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Design of modular autoproteolytic gene switches responsive to anti-coronavirus drug candidates"],"prefix":"10.1038","volume":"12","author":[{"given":"Nik","family":"Franko","sequence":"first","affiliation":[]},{"given":"Ana Palma","family":"Teixeira","sequence":"additional","affiliation":[]},{"given":"Shuai","family":"Xue","sequence":"additional","affiliation":[]},{"given":"Ghislaine","family":"Charpin-El Hamri","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8545-667X","authenticated-orcid":false,"given":"Martin","family":"Fussenegger","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2021,11,22]]},"reference":[{"key":"27072_CR1","doi-asserted-by":"publisher","first-page":"536","DOI":"10.1038\/s41564-020-0695-z","volume":"5","author":"AE Gorbalenya","year":"2020","unstructured":"Gorbalenya, A. 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