FUNCTIONAL ANALYSIS OF ESSENTIAL C. ALBICANS GENES

白色念珠菌必需基因的功能分析

• 批准号: 8301146
• 负责人: AARON P MITCHELL
• 金额: $ 23.04万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8301146
• 关键词: Address; Alleles; American; Anti-Infective Agents; Bacterial DNA; Biological; Biological Assay; Candida albicans; Cell Wall; Cell surface; Collection; Communities; Data; Data Set; Development; Drug Delivery Systems; Drug Industry; Essential Genes; Gene Expression; Genes; Genetic; Genetic Research; Genetic Translation; Goals; Growth; Health; Health Care Costs; Infection; Insertion Mutation; KRP protein; Knowledge; Measurement; Messenger RNA; Methods; Nuclear Export; Pathway interactions; Pharmaceutical Preparations; Pilot Projects; Poly-A Addition Site; Protein Kinase; Publishing; Regulatory Pathway; Reporter Genes; Research; Resources; Saccharomyces cerevisiae; Source; Therapeutic; acronyms; base; case-by-case basis; gene function; microbial; mutant; pathogen; prospective; success; therapeutic target; tool

DESCRIPTION (provided by applicant): Our focus is Candida albicans, the predominant invasive fungal pathogen in the USA. Our goal is to develop a genetic tool and a resource of strains and data that facilitate understanding of essential C. albicans genes, hence prospective therapeutic targets. Our studies rely upon a systematic approach to create partially defective alleles, called "DAmP" alleles ("decreased abundance by mRNA perturbation"), that we have borrowed from S. cerevisiae geneticists. DAmP alleles are created by replacing the 3' noncoding sequence of a gene with sequences that lack efficient cleavage and poly-A addition sites, thus reducing nuclear export, stability, and translation of the mRNA. In pilot studies we have applied the approach successfully to the essential C. albicans genes CDC3, CDC12, IRE1, SNF1, DBF2, and orf19.5376. We seek to extend the approach by implementing two specific aims: (1) to create DAmP strains for 17 protein kinase-related genes and 50 cell wall-related genes, all of which we have been unable to disrupt with insertion mutations; and (2) to assign these genes to functional pathways through DAmP strain phenotypic analysis. Completion of these aims will provide new understanding of C. albicans gene function, focused on a set of genes prioritized to include prospective therapeutic targets. In terms of deliverables, the project will yield new methodological tools, a characterized strain collection, and a dataset that will benefit the C. albicans research community. In terms of overall impact, we foresee that our success with this approach will promote its implementation more broadly in C. albicans and in other eukaryotic pathogens. PUBLIC HEALTH RELEVANCE: Fungal pathogens cause lethal infections in over 10,000 Americans per year and add over $2.6 billion to our health care costs. New drugs are needed to address this health threat. Our project will develop a new method to understand drug targets in fungal pathogens.

描述（由申请人提供）：我们的重点是白色念珠菌，它是美国主要的侵入性真菌病原体。我们的目标是开发一种遗传工具以及菌株和数据资源，以促进对白色念珠菌重要基因的理解，从而确定潜在的治疗靶点。我们的研究依赖于一种系统方法来创建部分有缺陷的等位基因，称为“DAmP”等位基因（“mRNA 扰动导致丰度减少”），这是我们从酿酒酵母遗传学家那里借来的。 DAmP 等位基因是通过用缺乏有效切割和多聚腺苷酸添加位点的序列替换基因的 3' 非编码序列来创建的，从而减少 mRNA 的核输出、稳定性和翻译。在试点研究中，我们已成功将该方法应用于白色念珠菌必需基因 CDC3、CDC12、IRE1、SNF1、DBF2 和 orf19.5376。我们寻求通过实现两个具体目标来扩展该方法：（1）为 17 个蛋白激酶相关基因和 50 个细胞壁相关基因创建 DAmP 菌株，所有这些基因我们都无法通过插入突变来破坏； (2) 通过 DAmP 菌株表型分析将这些基因分配给功能途径。这些目标的完成将为白色念珠菌基因功能提供新的认识，重点关注一组优先考虑前瞻性治疗靶点的基因。在可交付成果方面， 该项目将产生新的方法工具、特征菌株集合和数据集，这将有利于白色念珠菌研究界。就总体影响而言，我们预计这种方法的成功将促进其在白色念珠菌和其他真核病原体中更广泛的实施。 公共卫生相关性：真菌病原体每年导致超过 10,000 名美国人遭受致命感染，并导致我们的医疗保健费用增加超过 26 亿美元。需要新药来应对这一健康威胁。我们的项目将开发一种新方法来了解真菌病原体的药物靶点。