Pathway for functional characterization of hypothetical genes and non-coding RNAs of Enterococcus faecalis

粪肠球菌假设基因和非编码 RNA 功能表征途径

• 批准号: 8986937
• 负责人: GARY M DUNNY
• 金额: $ 22.8万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8986937
• 关键词: Antibiotic Resistance; Antibiotic Therapy; Biochemical; Biological Assay; Biology; Centers for Disease Control and Prevention (U.S.); Chemicals; Code; Computing Methodologies; Data; Databases; Disease; Endocarditis; Enterococcus faecalis; Environment; Experimental Models; Future; Genes; Genetic; Genetic Determinism; Genetic Screening; Genome; Growth; Hospitals; In Vitro; Infection; Intercistronic Region; Laboratories; Lead; Length; Life Style; Microarray Analysis; Microbial Biofilms; Modeling; Molecular; Molecular Mechanisms of Action; Mutation; Nosocomial Infections; Opportunistic Infections; Organism; Oryctolagus cuniculus; PAWR protein; Pathogenicity; Pathway interactions; Peptides; Pharmaceutical Preparations; Physiological; Physiology; Positioning Attribute; Production; Proteins; RNA; Regulation; Research; Resources; Role; Structure; Study models; System; Technology; Tertiary Protein Structure; Time; Untranslated RNA; Vaccines; antimicrobial drug; base; fitness; follow-up; genome sequencing; genome-wide; genome-wide analysis; in vivo; interest; mutant; novel; pathogen; polypeptide; protein function; public health relevance; recombinase; research study; transcriptome sequencing

 DESCRIPTION (provided by applicant): This project will examine the role of non-coding regulatory RNAs (ncRNAs) and hypothetical proteins of Enterococcus faecalis in adaptation to growth conditions relevant to opportunistic infections. E. faecalis is a leading cause of hospital-acquired infections whose treatment is impeded by the inherent and acquired antibiotic resistance of the organism, and by its propensity to adapt to growth and survival in hostile conditions, and by its ability to transition between commensal and pathogenic lifestyles. We will study the model laboratory strain OG1RF in both an in-vitro biofilm system (CDC biofilm reactor) and in an experimental endocarditis model. We will use a combination of computational searches, and a powerful new genetic screen we have termed "Smart TnSeq" to identify hypothetical proteins ncRNAs contributing to competitive fitness in these environments, and begin to study their targets and mechanisms of action. The results of this research will include the following: 1) A more comprehensive picture of the role of ncRNA regulation and hypothetical protein function in this organism under growth conditions relevant to its role in hospital infectios will emerge. 2) A workflow for genome-wide functional and mechanistic studies of regulation by E. faecalis ncRNAs and hypothetical proteins will be validated. 3) In the longer term, new drug and vaccine targets may be identified.

 描述（由申请人提供）：本项目将研究粪肠球菌的非编码调节RNA（ncRNA）和假设蛋白在适应与机会性感染相关的生长条件中的作用。E.粪菌是医院获得性感染的主要原因，其治疗受到生物体固有和获得性抗生素耐药性的阻碍，并受到其适应在恶劣条件下生长和生存的倾向以及其在寄生性和致病性生活方式之间转变的能力的阻碍。我们将在体外生物膜系统（CDC生物膜反应器）和实验性心内膜炎模型中研究模型实验室菌株OG1RF。我们将结合使用计算搜索和我们称为“智能TnSeq”的强大的新遗传筛选来识别在这些环境中有助于竞争适应性的假设蛋白质ncRNA，并开始研究它们的靶点和作用机制。本研究的结果将包括以下几点：1）在与其在医院感染中的作用相关的生长条件下，将出现一个更全面的ncRNA调节作用和假设蛋白质功能的图片。2)一个全基因组的功能和机制研究的工作流程调控E。将验证粪便ncRNA和假设蛋白质。3)从长远来看，可能会发现新的药物和疫苗靶点。