Essential Gene Discovery in the Basidiomycete Cryptococcus neoformans for Antifungal Drug Target Prioritization

摘要: ABSTRACT Fungal diseases represent a major burden to health care globally. As with other pathogenic microbes, there is limited number of agents suitable for use in treating fungal diseases, and resistance these can develop rapidly. Cryptococcus neoformans basidiomycete fungus that causes cryptococcosis worldwide both immunocompromised healthy individuals. basidiomycete, it diverged from common or model ascomycete fungi more than 500 million years ago. Here, we report C. genes are essential viability as identified through forward reverse genetic approaches, using an engineered diploid strain segregation after meiosis. The approach generated random insertional mutants the strain, induction meiosis sporulation, selection haploid cells counterselection insertion event. More 2,500 were analyzed, transfer DNA (T-DNA) insertions several required identified. include those encoding thioredoxin reductase (Trr1), ribosome assembly factor (Rsa4), mRNA-capping component (Cet1), others. For targeted gene replacement, homologs 35 disrupted, sporulation induced, progeny evaluated their ability grow on selective media. Twenty-one (60%) found be neoformans. These involved mitochondrial translation, ergosterol biosynthesis, RNA-related functions. heterozygous haploinsufficiency perturbing drugs, revealing phenotypes due loss one copy This study expands knowledge organism. Genes have no mammalian human pathogens would ideal development antifungal drugs broad-spectrum activity. IMPORTANCE infections very humans but may neglected misdiagnosis inattention. yeast infects mainly people, causing high mortality rates developing countries. lungs, crosses blood-brain barrier, invades cerebrospinal fluid, fatal meningitis. treated amphotericin B, flucytosine, azoles, all developed decades However, problems highlight urgent need more-effective treat invasive infections. issues negative side effects spontaneous inefficacy echinocandin antifungals. In this study, identification targets novel Because level evolutionary divergence between ascomycetes, subset likely fungi. excellent starting point future new antifungals by pharmaceutical companies.