{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:04:59Z","timestamp":1792785899495,"version":"4.5.0"},"reference-count":174,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T00:00:00Z","timestamp":1669852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NSERC Discovery","award":["RGPIN-2018-4914"],"award-info":[{"award-number":["RGPIN-2018-4914"]}]},{"name":"Ontario Graduate Scholarship"},{"name":"MITACS Globalink Research Internship"},{"name":"Medical Scientist"},{"DOI":"10.13039\/100000861","name":"Burroughs Wellcome Fund","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000861","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Institute of Health","award":["R01-DK128805"],"award-info":[{"award-number":["R01-DK128805"]}]},{"name":"Schmitt Presidential Leadership Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,2,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>For the fungal pathogen Candida albicans, genetic overexpression readily occurs via a diversity of genomic alterations, such as aneuploidy and gain-of-function mutations, with important consequences for host adaptation, virulence, and evolution of antifungal drug resistance. Given the important role of overexpression on C. albicans biology, it is critical to develop and harness tools that enable the analysis of genes expressed at high levels in the fungal cell. Here, we describe the development, optimization, and application of a novel, single-plasmid-based CRISPR activation (CRISPRa) platform for targeted genetic overexpression in C. albicans, which employs a guide RNA to target an activator complex to the promoter region of a gene of interest, thus driving transcriptional expression of that gene. Using this system, we demonstrate the ability of CRISPRa to drive high levels of gene expression in C. albicans, and we assess optimal guide RNA targeting for robust and constitutive overexpression. We further demonstrate the specificity of the system via RNA sequencing. We highlight the application of CRISPR activation to overexpress genes involved in pathogenesis and drug susceptibility, and contribute toward the identification of novel phenotypes. Consequently, this tool will facilitate a broad range of applications for the study of C. albicans genetic overexpression.<\/jats:p>","DOI":"10.1093\/g3journal\/jkac301","type":"journal-article","created":{"date-parts":[[2022,11,30]],"date-time":"2022-11-30T20:53:17Z","timestamp":1669841597000},"source":"Crossref","is-referenced-by-count":32,"title":["Development and applications of a CRISPR activation system for facile genetic overexpression in\n                    <i>Candida albicans<\/i>"],"prefix":"10.1093","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9971-0031","authenticated-orcid":false,"given":"Nicholas C","family":"Gervais","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alyssa A","family":"La Bella","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Notre Dame , Notre Dame, IN 46556 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren F","family":"Wensing","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jehoshua","family":"Sharma","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Acquaviva","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madison","family":"Best","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ricardo Omar","family":"Cadena L\u00f3pez","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , 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, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deeva","family":"Uthayakumar","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Biology, University of Guelph , Guelph, ON N1H 5N4 , Canada"},{"name":"Present address: Department of Immunology, University of Toronto , Toronto, ON , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro","family":"Chavez","sequence":"additional","affiliation":[{"name":"Department of Pathology and Cell Biology, Columbia University College of Physicians and Surgeons , New York, NY 10032 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felipe","family":"Santiago-Tirado","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Notre Dame , Notre Dame, IN 46556 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana L","family":"Flores-Mireles","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Notre Dame , Notre Dame, IN 46556 , USA"}],"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, ON N1H 5N4 , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,12,1]]},"reference":[{"issue":"1","key":"2024022107385302300_jkac301-B1","doi-asserted-by":"publisher","first-page":"1911","DOI":"10.1038\/s41467-018-04252-2","article-title":"The CRISPR tool kit for genome editing and beyond","volume":"9","author":"Adli","year":"2018","journal-title":"Nat Commun."},{"issue":"1","key":"2024022107385302300_jkac301-B2","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/s40694-018-0057-2","article-title":"Whole-genome sequencing reveals highly specific gene targeting by in vitro assembled Cas9-ribonucleoprotein complexes in Aspergillus fumigatus","volume":"5","author":"Al Abdallah","year":"2018","journal-title":"Fungal Biol 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