{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T07:15:28Z","timestamp":1791357328790,"version":"4.3.1"},"reference-count":69,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2015,3,31]],"date-time":"2015-03-31T00:00:00Z","timestamp":1427760000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-sa\/3.0\/"},{"start":{"date-parts":[[2015,5,1]],"date-time":"2015-05-01T00:00:00Z","timestamp":1430438400000},"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":["mBio"],"published-print":{"date-parts":[[2015,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Fungal diseases represent a major burden to health care globally. As with other pathogenic microbes, there is a limited number of agents suitable for use in treating fungal diseases, and resistance to these agents can develop rapidly.\n                    <jats:named-content content-type=\"genus-species\">Cryptococcus neoformans<\/jats:named-content>\n                    is a basidiomycete fungus that causes cryptococcosis worldwide in both immunocompromised and healthy individuals. As a basidiomycete, it diverged from other common pathogenic or model ascomycete fungi more than 500 million\u00a0years ago. Here, we report\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    genes that are essential for viability as identified through forward and reverse genetic approaches, using an engineered diploid strain and genetic segregation after meiosis. The forward genetic approach generated random insertional mutants in the diploid strain, the induction of meiosis and sporulation, and selection for haploid cells with counterselection of the insertion event. More than 2,500 mutants were analyzed, and transfer DNA (T-DNA) insertions in several genes required for viability were identified. The genes include those encoding the thioredoxin reductase (Trr1), a ribosome assembly factor (Rsa4), an mRNA-capping component (Cet1), and others. For targeted gene replacement, the\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    homologs of 35 genes required for viability in ascomycete fungi were disrupted, meiosis and sporulation were induced, and haploid progeny were evaluated for their ability to grow on selective media. Twenty-one (60%) were found to be required for viability in\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    . These genes are involved in mitochondrial translation, ergosterol biosynthesis, and RNA-related functions. The heterozygous diploid mutants were evaluated for haploinsufficiency on a number of perturbing agents and drugs, revealing phenotypes due to the loss of one copy of an essential gene in\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    . This study expands the knowledge of the essential genes in fungi using a basidiomycete as a model organism. Genes that have no mammalian homologs and are essential in both\n                    <jats:italic>Cryptococcus<\/jats:italic>\n                    and ascomycete human pathogens would be ideal for the development of antifungal drugs with broad-spectrum activity.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>IMPORTANCE<\/jats:bold>\n                    Fungal infections are very common in humans but may be neglected due to misdiagnosis and inattention.\n                    <jats:named-content content-type=\"genus-species\">Cryptococcus neoformans<\/jats:named-content>\n                    is a yeast that infects mainly immunocompromised people, causing high mortality rates in developing countries. The fungus infects the lungs, crosses the blood-brain barrier, and invades the cerebrospinal fluid, causing fatal meningitis.\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    infections are treated with amphotericin B, flucytosine, and azoles, all developed decades ago. However, problems with antifungal agents highlight the urgent need for more-effective drugs to treat\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    and other invasive fungal infections. These issues include the negative side effects of amphotericin B, the spontaneous resistance of\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    to azoles, and the inefficacy of the echinocandin antifungals. In this study, we report the identification of\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    essential genes as targets for the development of novel antifungals. Because of the level of evolutionary divergence between\n                    <jats:named-content content-type=\"genus-species\">C. neoformans<\/jats:named-content>\n                    and the ascomycetes, a subset of these genes is likely essential in all fungi. Genes identified in this study represent an excellent starting point for the future development of new antifungals by pharmaceutical companies.\n                  <\/jats:p>","DOI":"10.1128\/mbio.02334-14","type":"journal-article","created":{"date-parts":[[2015,4,1]],"date-time":"2015-04-01T08:47:31Z","timestamp":1427878051000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":60,"title":["Essential Gene Discovery in the Basidiomycete Cryptococcus neoformans for Antifungal Drug Target Prioritization"],"prefix":"10.1128","volume":"6","author":[{"given":"Giuseppe","family":"Ianiri","sequence":"first","affiliation":[{"name":"Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri\u2014Kansas City, Missouri, USA"},{"name":"Dipartimento di Agricoltura, Ambiente e Alimenti, Universit\u00e0 del Molise, Campobasso, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Idnurm","sequence":"additional","affiliation":[{"name":"Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri\u2014Kansas City, Missouri, USA"},{"name":"School of Biosciences, University of Melbourne, Melbourne, Victoria, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.3004404"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/9781555818241"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/9781555816858"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1524-4733.2002.51108.x"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/myc.12238"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1097\/QAD.0b013e328322ffac"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1508.081384"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000850"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0074737"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000848"},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","unstructured":"Mondon P Petter R Amalfitano G Luzzati R Concia E Polacheck I Kwon-Chung KJ. 1999. 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