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Here we systematically investigate essential transcriptional regulators in\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  as potential antifungal targets, developing experimental pipelines that assess growth requirement, essentiality and function through conditional gene expression, constitutive overexpression, and meiotic spore analysis. We identify one quasi-essential (growth-required but non-essential) transcription factor, Fhl1, and 13 essential transcriptional regulators, three of which are transcription factors (Ezt1, Ezt2 and Cbf1) highly divergent from counterparts in other eukaryotes. Notably, Ezt1 modulates the expression of more than 1,200 genes, controlling growth, antifungal drug and stress responses, sexual development and virulence. Our findings define the essential transcriptional regulator landscape of\n                  <jats:italic>C. neoformans<\/jats:italic>\n                  and provide a framework for prioritising divergent essential regulators, particularly Ezt1, for antifungal target discovery.\n                <\/jats:p>","DOI":"10.64898\/2026.04.30.722081","type":"posted-content","created":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T20:50:16Z","timestamp":1778014216000},"source":"Crossref","is-referenced-by-count":0,"title":["Systematic Profiling of Essential Fungal Transcriptional Regulators Uncovers Ezt1 as a Central Pathobiological and Morphogenic Regulator in\n                  <i>Cryptococcus neoformans<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0578-7653","authenticated-orcid":false,"given":"Seung-Heon","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea"},{"name":"Department of Molecular Genetics, University of Toronto, Toronto, Ontario, M5G 1M1, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2895-1153","authenticated-orcid":false,"given":"Sheng","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5100-7654","authenticated-orcid":false,"given":"Yu-Byeong","family":"Jang","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0025-3860","authenticated-orcid":false,"given":"Seong-Ryong","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9369-902X","authenticated-orcid":false,"given":"Yeseul","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3077-1116","authenticated-orcid":false,"given":"Jin-Tae","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-7836-4583","authenticated-orcid":false,"given":"Eui-Seong","family":"Kim","sequence":"additional","affiliation":[{"name":"Korea Zoonosis Research Institute, Jeonbuk National University, Iksan, Jeonbuk State 54531, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6257-331X","authenticated-orcid":false,"given":"Jong-Seung","family":"Lee","sequence":"additional","affiliation":[{"name":"AmtixBio Co., Ltd., Seoul 05836, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5797-0110","authenticated-orcid":false,"given":"Leah E.","family":"Cowen","sequence":"additional","affiliation":[{"name":"Department of Molecular Genetics, University of Toronto, Toronto, Ontario, M5G 1M1, 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