Cryptococcus neoformans Gene Knockout Resource

新型隐球菌基因敲除资源

基本信息

批准号：
9052114
负责人：
Hiten D Madhani
金额：
$ 51.85万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9052114
关键词：
AIDS/HIV problem Accounting Acquired Immunodeficiency Syndrome Address Affect Bar Codes Biology Breathing Cell Wall Cessation of life Cities Clinical Collection Communities Cryptococcus neoformans Data Data Set Databases Defect Development Diagnosis Discipline Disease Encapsulated Environment Essential Genes Fission Yeast Foundations Fungal Meningitis Fungi Model Gene Deletion Gene Targeting Generations Genes Genomics Human Hypoxia In Vitro Individual Infection Injection of therapeutic agent Iron Kansas Knock-out Laboratories Letters Libraries Life Link Lung Medical Melanins Methods Missouri Modeling Modification Mus Mycoses Organism Patients Phagocytosis Phagocytosis Inhibition Pharmaceutical Preparations Phenotype Polysaccharides Production Property Publishing Reporting Research Resistance Resources Saccharomyces cerevisiae Saccharomycetales Series Stress The science of Mycology Therapeutic Universities Urease Validation Virulence Virulence Factors Work base capsule deletion library design experience fitness functional genomics fungal genetics fungus genetic strain genome-wide in vitro Assay in vivo knockout gene mutant nitrosative stress particle pathogen relational database research study time use tool trait web-accessible whole genome

项目摘要

DESCRIPTION (provided by applicant): The effective diagnosis and treatment of life-threatening fungal infections in humans is a major unmet clinical challenge, and medical mycology is the research discipline that addresses this need. Well-annotated genomic sequences of the major human fungal pathogens have recently been published, creating an opportunity to revolutionize medical mycology through the application of systematic genome-wide approaches. Just as the availability of genome-wide knockout collections has been instrumental in dissecting the biology of model fungi such as Saccharomyces cerevisiae and Schizosaccharomyces pombe, the generation of analogous complete gene deletion collections in pathogenic fungi would create experimental tools to systematically elucidate the basis of fungal virulence in the mammalian host and to develop drugs against them. Cryptococcus neoformans is an encapsulated budding yeast that is the most common cause of fungal meningitis. There are an estimated 1,000,000 cases that result in ~600,000 deaths each year. It is estimated to cause one-third of deaths in AIDS patients worldwide. Using optimized particle bombardment methods for gene targeting, our laboratory demonstrated the feasibility of creating large numbers of bar-coded gene deletions in this organism. Specifically, we reported previously the construction of 1201 gene deletion strains. This collection has been made available without restriction to the community. We demonstrated the utility of this collection by using it in systematic screens of infectivity in experimental mice, expression of known virulence factors, mechanisms of hypoxic adaptation, and mechanisms of phagocytosis-inhibition. However, the impact of this collection is limited to the fraction of genes covered -- one-fifth of predicted gens. Objectives: We propose to generate and distribute a complete collection of knockout mutants for Cryptococcus neoformans. Knockouts will contain features to facilitate functional genomic studies. We further propose to obtain a series of reference phenotypes via quantitative screens of the library for defects in infectivity in vivo and the expression of the major virulence factorsin vitro. The proposed efort will be coordinated with those of others that aim to identify the essential genes of C. neoformans. Impact: The proposed work will transform medical mycology by providing the first full description of pathogen genes required for fitness during infection, a ull accounting of genes required for production of known virulence factors, and a permanent freely-available, nonredundant null mutant strain resource that will dramatically accelerate research in the field as a whole.

描述（由申请人提供）：对人类危及生命的真菌感染的有效诊断和治疗是一个重大的未满足的临床挑战，而医学真菌学是解决这一需求的研究学科。最近发表了详细注释的人类主要真菌病原体的基因组序列，为通过应用系统的全基因组方法彻底改变医学真菌学创造了机会。正如全基因组敲除集合的可用性有助于剖析酿酒酵母和粟酒裂殖酵母等模式真菌的生物学一样，致病性真菌中类似的完整基因删除集合的产生也将创建实验工具来系统地阐明真菌的基础。哺乳动物宿主的毒力并开发针对它们的药物。新型隐球菌是一种封装的芽殖酵母，是真菌性脑膜炎的最常见原因。据估计，每年有 1,000,000 例病例导致约 600,000 人死亡。据估计，全世界三分之一的艾滋病患者死亡是由它造成的。我们的实验室使用优化的粒子轰击方法进行基因靶向，证明了在该生物体中产生大量条形码基因缺失的可行性。具体来说，我们之前报道了1201个基因缺失菌株的构建。该集合已不受限制地向社区开放。我们通过将其用于实验小鼠感染性的系统筛选、已知毒力因子的表达、缺氧适应机制和吞噬抑制机制来证明该集合的实用性。然而，这个集合的影响仅限于所覆盖的基因部分——预测基因的五分之一。目标：我们建议生成并分发完整的新型隐球菌敲除突变体集合。基因敲除将包含促进功能基因组研究的功能。我们进一步建议通过定量筛选文库的体内感染性缺陷和体外主要毒力因子的表达来获得一系列参考表型。拟议的工作将与其他旨在识别新型隐球菌必需基因的工作相协调。影响：拟议的工作将通过提供对感染过程中健康所需的病原体基因的首次完整描述、对产生已知毒力因子所需的基因的完整说明以及永久免费提供的非冗余无效突变株资源来改变医学真菌学。将极大地加速整个领域的研究。