{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T21:00:25Z","timestamp":1790283625558,"version":"4.1.0"},"reference-count":50,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/doi.org\/10.1128\/ASMCopyrightv2"},{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              <jats:italic>Candida parapsilosis<\/jats:italic>\n              has recently emerged as a major threat due to the worldwide emergence of fluconazole-resistant strains causing clonal outbreaks in hospitals and poses a therapeutic challenge due to the limited antifungal armamentarium. Here, we used precise genome editing using CRISPR-Cas9 to gain further insights into the contribution of mutations in\n              <jats:italic>ERG11<\/jats:italic>\n              ,\n              <jats:italic>ERG3<\/jats:italic>\n              ,\n              <jats:italic>MRR1<\/jats:italic>\n              , and\n              <jats:italic>TAC1<\/jats:italic>\n              genes and the influence of allelic dosage to antifungal resistance in\n              <jats:italic>C. parapsilosis<\/jats:italic>\n              . Seven of the most common amino acid substitutions previously reported in fluconazole-resistant clinical isolates (including Y132F in\n              <jats:italic>ERG11<\/jats:italic>\n              ) were engineered in two fluconazole-susceptible\n              <jats:italic>C. parapsilosis<\/jats:italic>\n              lineages (ATCC 22019 and STZ5). Each mutant was then challenged\n              <jats:italic>in vitro<\/jats:italic>\n              against a large array of antifungals, with a focus on azoles. Any possible change in virulence was also assessed in a\n              <jats:italic>Galleria mellonella<\/jats:italic>\n              model. We successfully generated a total of 19 different mutants, using CRISPR-Cas9. Except for R398I (\n              <jats:italic>ERG11<\/jats:italic>\n              ), all remaining amino acid substitutions conferred reduced susceptibility to fluconazole. However, the impact on fluconazole\n              <jats:italic>in vitro<\/jats:italic>\n              susceptibility varied greatly according to the engineered mutation, the stronger impact being noted for G583R acting as a gain-of-function mutation in\n              <jats:italic>MRR1<\/jats:italic>\n              . Cross-resistance with newer azoles, non-medical azoles, but also non-azole antifungals such as flucytosine, was occasionally noted. Posaconazole and isavuconazole remained the most active\n              <jats:italic>in vitro<\/jats:italic>\n              . Except for G583R, no fitness cost was associated with the acquisition of fluconazole resistance. We highlight the distinct contributions of amino acid substitutions in\n              <jats:italic>ERG11<\/jats:italic>\n              ,\n              <jats:italic>ERG3<\/jats:italic>\n              ,\n              <jats:italic>MRR1<\/jats:italic>\n              , and\n              <jats:italic>TAC1<\/jats:italic>\n              genes to antifungal resistance in\n              <jats:italic>C. parapsilosis<\/jats:italic>\n              .\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/aac.00022-24","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T13:09:35Z","timestamp":1713272975000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Precise genome editing underlines the distinct contributions of mutations in\n            <i>ERG11<\/i>\n            ,\n            <i>ERG3<\/i>\n            ,\n            <i>MRR1<\/i>\n            , and\n            <i>TAC1<\/i>\n            genes to antifungal resistance in\n            <i>Candida parapsilosis<\/i>"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-4284-8802","authenticated-orcid":false,"given":"Sophie","family":"Hartuis","sequence":"first","affiliation":[{"name":"Nantes Universit\u00e9, CHU Nantes, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabelle","family":"Ourliac-Garnier","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estelle","family":"Robert","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marjorie","family":"Albassier","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L\u00e9a","family":"Duchesne","sequence":"additional","affiliation":[{"name":"Department Public Health, Nantes Universit\u00e9, CHU Nantes, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clara","family":"Beaufils","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9phine","family":"Kuhn","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Le Pape","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, CHU Nantes, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5859-6105","authenticated-orcid":true,"given":"Florent","family":"Morio","sequence":"additional","affiliation":[{"name":"Nantes Universit\u00e9, CHU Nantes, Cibles et M\u00e9dicaments des Infections et de l'Immunit\u00e9, Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"key":"e_1_3_4_2_2","unstructured":"World Health Organization. 2022. 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