{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,12]],"date-time":"2026-04-12T19:29:15Z","timestamp":1776022155097,"version":"3.50.1"},"posted":{"date-parts":[[2025,2,11]]},"group-title":"Microbiology","reference-count":30,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T00:00:00Z","timestamp":1739232000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,2,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Objective<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Candida auris<\/jats:italic>\n                    has emerged as a fungal pathogen of particular concern owing in part to its propensity to exhibit antifungal resistance, especially to the commonly prescribed antifungal fluconazole. In this work we aimed to determine how mutations in the transcription factor gene\n                    <jats:italic>TAC1B<\/jats:italic>\n                    , which are common among resistant isolates and confer fluconazole resistance, exert this effect.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    Selected\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations from clinical isolates were introduced into a susceptible isolate and reverted to the wild-type sequence in select clinical isolates using CRISPR Cas9 gene editing. Disruption mutants were likewise generated for select genes of interest.\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutants were subjected to transcriptional profiling by RNA-seq, and relative expression of specific genes of interest was determined by qRT-PCR. Antifungal susceptibilities were determined by modified CLSI broth microdilution.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations leading to A640V, A657V, and F862_N866del conferred fluconazole resistance, as well as increased resistance to other triazoles, when introduced into a susceptible isolate. RNA-seq revealed that the ATP-Binding Cassette (ABC) transporter gene\n                    <jats:italic>CDR1<\/jats:italic>\n                    as well as the Major Facilitator Superfamily (MFS) transporter gene\n                    <jats:italic>MDR1<\/jats:italic>\n                    were both upregulated by these\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations. Disruption of\n                    <jats:italic>CDR1<\/jats:italic>\n                    greatly abrogated resistance in strains with\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations whereas disruption of\n                    <jats:italic>MDR1<\/jats:italic>\n                    had little to no effect. However, disruption of both\n                    <jats:italic>CDR1<\/jats:italic>\n                    and\n                    <jats:italic>MDR1<\/jats:italic>\n                    resulted in an additional reduction in resistance as compared to disruption of either gene alone.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations leading to A640V, A657V, and F862_N866del all result in increased resistance to fluconazole and other triazole antifungals, and increased expression of both\n                    <jats:italic>CDR1<\/jats:italic>\n                    and\n                    <jats:italic>MDR1<\/jats:italic>\n                    in\n                    <jats:italic>C. auris<\/jats:italic>\n                    .\n                    <jats:italic>CDR1<\/jats:italic>\n                    is the primary driver of resistance conferred by these\n                    <jats:italic>TAC1B<\/jats:italic>\n                    mutations.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.02.11.637698","type":"posted-content","created":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T21:45:38Z","timestamp":1739310338000},"source":"Crossref","is-referenced-by-count":5,"title":["Mutations in\n                  <i>TAC1B<\/i>\n                  drive\n                  <i>CDR1<\/i>\n                  and\n                  <i>MDR1<\/i>\n                  expression and azole resistance in\n                  <i>C. auris<\/i>"],"prefix":"10.64898","author":[{"given":"Katherine S.","family":"Barker","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Darian J.","family":"Santana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qing","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tracy L.","family":"Peters","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey M.","family":"Rybak","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joachim","family":"Morschh\u00e4user","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christina A.","family":"Cuomo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P. 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