Silencing of Mobile Genetic Elements by Small RNAs

小RNA沉默移动遗传元件

• 批准号: 8565082
• 负责人: Carolyn Marie Phillips
• 金额: $ 19.17万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8565082
• 关键词: Affect; Biochemistry; Biological Assay; Biology; Caenorhabditis elegans; Chromatin; Chromatin Structure; Commit; Congenital Abnormality; Critiques; DNA; DNA Damage; DNA Transposons; Data; Development; Disease; Environment; Enzymes; Functional RNA; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Screening; Genome; Genomic Instability; Genomics; Goals; High-Throughput Nucleotide Sequencing; Housing; In Vitro; Individual; Infertility; Laboratories; Lead; Malignant Neoplasms; Methods; Mobile Genetic Elements; Movement; Mutation; Neurobiology; Pathogenesis; Pathway interactions; Phosphodiesterase I; Positioning Attribute; Postdoctoral Fellow; Process; Proteins; RNA; RNA Interference; Reagent; Regulation; Research; Research Personnel; Resources; Rest; Retrotransposon; Role; Small RNA; Stimulus; Stress; Structure; Substrate Specificity; Surveys; Therapeutic; Transcript; Work; Writing; biological adaptation to stress; cancer diagnosis; cancer prevention; career; genome-wide; in vivo; interest; meetings; mutant; novel; prevent; protein complex; protein purification; public health relevance; research study; skills training; tool

DESCRIPTION (provided by applicant): The goal of this project is to characterize the transposon-silencing pathway, specifically focusing on the endogenous role of small RNAs. Since the late 1980's, transposons have been well characterized in C. elegans through the use of classical genetic tools to interrogate questions related to mechanisms of transposition. In 1999, it was discovered that endogenous small RNA pathways regulate transposons and several screens were performed to isolate mutants with active transposition in the germline. However, although many mutants have been isolated in genes attributed to this pathway, little work has been done to mechanistically characterize the roles of individual proteins. Additionally, because most analyses of transposon movement were done prior to the advent of high-throughput sequencing, there is no genome-wide data examining transposon mobilization. In addition to transposons, many other types of transcripts are processed through the endogenous RNA pathways, including aberrant transcripts, duplicated and hypothetical genes, and non-coding regions, however the mechanism by which these transcripts are recognized and routed into these pathways is completely unknown. By probing the individual components of these pathways, examining what species of RNA they associate with, and to what process their enzymatic functions contribute, this project will achieve a more complete understanding of transposon silencing. The proposed experiments herein aim to identify factors that regulate transposon mobilization, including genomic features, host proteins, and environmental stimuli with the goal of elucidating the mechanisms of action of the known and novel components of this pathway. The results of this work will have significant implications in understanding how active transposons affect gene expression and alter genome structure, which can result in many diseases including cancer. The Ruvkun lab is an ideal place to study regulation of transposons by RNAi machinery in C. elegans. The lab not only has extensive expertise in the small RNA field, but also in stress response pathways, high- throughput genetic screens, protein purification, and biochemistry. The laboratory environment contains an extremely motivated and scientifically diverse group of researchers interested in fundamental aspects of C. elegans biology. The lab is flanked by two excellent C. elegans labs studying neurobiology and pathogenesis and is supported by a highly collaborative department that houses labs with expertise in RNA biology and chromatin structure. I plan to utilize all of these resources during the rest of my post-doctoral tenure. Throughout my research career, I have been focused on chromatin and RNA biology, which are fundamentally important in the understanding and prevention of cancer. Specifically, greater understanding of RNA silencing and transposon quiescence may lead to advances in cancer diagnoses or therapeutics. I am committed to a career in academic research and plan to start a lab focused on the aims in this proposal.

描述（由申请人提供）：该项目的目标是表征转座子沉默途径，特别关注小RNA的内源作用。自 20 世纪 80 年代末以来，通过使用经典遗传工具来探究与转座机制相关的问题，人们已经对秀丽隐杆线虫中的转座子进行了很好的表征。 1999年，人们发现内源小RNA途径调节转座子，并进行了多次筛选以分离出种系中具有活性转座的突变体。然而，尽管许多突变体已在归因于该途径的基因中被分离出来，但在机械地表征单个蛋白质的作用方面几乎没有做任何工作。此外，由于大多数转座子运动分析是在高通量测序出现之前完成的，因此没有检查转座子动员的全基因组数据。 除了转座子之外，许多其他类型的转录本也通过内源 RNA 途径进行加工，包括异常转录本、重复和假设的基因以及非编码区，但是这些转录本被识别并进入这些途径的机制完全未知。通过探测这些途径的各个组成部分，检查它们与哪些种类的 RNA 相关，以及它们的酶功能对什么过程做出贡献，该项目将实现对转座子沉默的更全面的了解。本文提出的实验旨在确定调节转座子动员的因素，包括基因组特征、宿主蛋白和环境刺激，目的是阐明该途径的已知和新成分的作用机制。这项工作的结果将对理解活性转座子如何影响基因表达和改变基因组结构产生重大影响，这可能导致包括癌症在内的许多疾病。 Ruvkun 实验室是研究秀丽隐杆线虫中 RNAi 机制对转座子调节的理想场所。该实验室不仅在小RNA领域拥有丰富的专业知识，而且在应激反应途径、高通量遗传筛选、蛋白质纯化和生物化学方面也拥有丰富的专业知识。实验室环境包含一群积极主动且科学多样化的研究人员，他们对秀丽隐杆线虫生物学的基本方面感兴趣。该实验室两侧是两个研究神经生物学和发病机制的优秀线虫实验室，并得到高度协作的部门的支持，该部门拥有在 RNA 生物学和染色质结构方面拥有专业知识的实验室。我计划在我剩余的博士后任期内利用所有这些资源。 在我的研究生涯中，我一直专注于染色质和 RNA 生物学，这对于理解和预防癌症至关重要。具体来说，对 RNA 沉默和转座子静止的更多了解可能会促进癌症诊断或治疗的进步。我致力于学术研究事业，并计划建立一个专注于本提案中目标的实验室。