Cryptococcus neoformans Gene Knockout Resource

新型隐球菌基因敲除资源

基本信息

批准号：
8836946
负责人：
Hiten D Madhani
金额：
$ 66.13万
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 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8836946
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 Genome 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

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）：在人类中有效诊断和治疗威胁生命的真菌感染是一个主要的临床挑战，医学真菌学是解决这一需求的研究学科。最近发表的主要人类真菌病原体的基因组序列被宣布良好，创造了通过应用全面基因组方法彻底改变医学真菌学的机会。正如全基因组淘汰集合的可用性在解剖诸如酿酒酵母和sacchizosacachomyces pombe的模型生物学方面发挥了作用一样，病原性真菌中的类似完整的基因缺失收集产生，可以使实验工具产生实验性的工具，从而有系统地依靠unt型养育型药物，并在妈妈中脱颖而出。加密赛车是一种封装的萌芽酵母菌，是真菌脑膜炎的最常见原因。估计有1,000,000例每年造成约600,000例死亡。据估计，它在全球艾滋病患者中造成三分之一的死亡。我们的实验室使用优化的颗粒轰击方法来靶向基因靶向，证明了在该生物体中产生大量条形码基因缺失的可行性。具体而言，我们先前报道了1201个基因缺失菌株的构建。该集合已不限于社区提供。我们通过在实验小鼠的系统感染性筛查中使用该收集的实用性，表达已知的毒力因子，低氧适应性机制以及抑制吞噬作用的机制。但是，该集合的影响仅限于涵盖基因的比例 - 占预测的五分之一。目的：我们建议生成和分发用于加密型新型人体的敲除突变体的完整集合。敲除将包含促进功能基因组研究的特征。我们进一步建议通过文库的定量筛选获得一系列参考表型，以使体内感染性缺陷以及体外主要毒力因子的表达。拟议的EFORT将与旨在识别新梭菌基本基因的其他人协调。影响：拟议的工作将通过提供对感染过程中适应性所需的病原体基因的第一个完整描述，对生产已知毒力因子产生所需的基因以及永久性的，无冗余的零突变菌株所需的基因来改变医学真菌学，该基因将在整个领域都大大加速研究。