{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T00:47:19Z","timestamp":1789519639360,"version":"build-2803163510"},"posted":{"date-parts":[[2025,12,22]]},"group-title":"Microbiology","reference-count":66,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,22]],"date-time":"2025-12-22T00:00:00Z","timestamp":1766361600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"crossref","award":["PJT 162195"],"award-info":[{"award-number":["PJT 162195"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["U19 AI110818"],"award-info":[{"award-number":["U19 AI110818"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The rising rate of drug-resistant fungal infections and the emergence of fungal pathogens with intrinsic resistance phenotypes are a growing concern. The close evolutionary distance between mammals and fungi complicates the design of new antifungals and increases the chances of toxic off-target effects. As such, antifungal drug development usually focuses on fungal-specific proteins when considering potential new targets. Ideal drug targets should mediate essential cell processes and be highly sensitive to inhibition. Targeted gene repression can serve as a model for drug-mediated inhibition and for determining the dosage-sensitivity profile of genes of interest. In the fungal pathogen\n                  <jats:italic>Candida albicans<\/jats:italic>\n                  , classical approaches for gene repression can be labour-intensive and limited to one genetic background due to low throughput. Here, we adapt pooled CRISPRi screening in\n                  <jats:italic>C. albicans<\/jats:italic>\n                  for the first time and exploit this technique for large-scale functional genomic analysis. Through pooled CRISPRi screening, we test the repression sensitivity of over a hundred essential genes conserved in fungi but absent in humans, and successfully identify highly dosage-sensitive genes across multiple cell components and pathways. By extending our analysis to ten diverse environmental conditions, we show how the environment influences dosage-sensitivity profiles. Finally, we extend our experiments to two clinical drug-resistant\n                  <jats:italic>C. albicans<\/jats:italic>\n                  strain backgrounds and demonstrate that many of the fitness defects we observed are conserved in resistant clinical isolates. Together, our results highlight a set of genes that are highly dosage-sensitive across different genetic and environmental contexts, making them attractive targets for further investigation. By facilitating rapid, efficient large-scale functional genomics assays across diverse genetic backgrounds, CRISPRi pooled screening will open new frontiers in\n                  <jats:italic>C. albicans<\/jats:italic>\n                  biology.\n                <\/jats:p>","DOI":"10.64898\/2025.12.19.695556","type":"posted-content","created":{"date-parts":[[2025,12,23]],"date-time":"2025-12-23T03:00:11Z","timestamp":1766458811000},"source":"Crossref","is-referenced-by-count":1,"title":["Pooled CRISPRi screening reveals fungal-specific vulnerabilities across environments and genetic backgrounds"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3737-5474","authenticated-orcid":false,"given":"Lauren F","family":"Wensing","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1407-0643","authenticated-orcid":false,"given":"Philippe C","family":"Despr\u00e9s","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"},{"name":"Infectious Disease and Microbiome Program, Broad Institute, Boston, United States"},{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-9003-5043","authenticated-orcid":false,"given":"Desiree","family":"Francis","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0483-9626","authenticated-orcid":false,"given":"Meea","family":"Fogal","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3679-4029","authenticated-orcid":false,"given":"Anthony","family":"Hendriks","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9971-0031","authenticated-orcid":false,"given":"Nicholas C","family":"Gervais","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3016-2314","authenticated-orcid":false,"given":"Clara","family":"Fikry","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0923-3893","authenticated-orcid":false,"given":"Abdul-Rahman","family":"Adamu Bukari","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, University of Manitoba, Winnipeg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0781-9356","authenticated-orcid":false,"given":"Aleeza C","family":"Gerstein","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Science, University of Manitoba, Winnipeg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5778-960X","authenticated-orcid":false,"given":"Christina A","family":"Cuomo","sequence":"additional","affiliation":[{"name":"Infectious Disease and Microbiome Program, Broad Institute, Boston, United States"},{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7119-8865","authenticated-orcid":false,"given":"Rebecca S","family":"Shapiro","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph, Guelph, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026011004151033000_2025.12.19.695556v2.1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-022-00720-1"},{"key":"2026011004151033000_2025.12.19.695556v2.2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(23)00692-8"},{"key":"2026011004151033000_2025.12.19.695556v2.3","doi-asserted-by":"crossref","first-page":"2338","DOI":"10.1038\/s41564-025-02084-7","article-title":"FungAMR: a comprehensive database for investigating fungal mutations associated with antimicrobial resistance","volume":"10","year":"2025","journal-title":"Nat. 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