Molecular Mechanisms of microRNA and miRISC turnover

microRNA 和 miRISC 更新的分子机制

• 批准号: 9115651
• 负责人: Katherine McJunkin
• 金额: $ 7.33万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-03-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9115651
• 关键词: 3' Untranslated Regions; Acute; Advisory Committees; Animals; Binding; Biochemistry; Biogenesis; Biological Models; Biology; Boxing; CRISPR/Cas technology; Caenorhabditis elegans; Candidate Disease Gene; Cellular biology; Cleaved cell; Code; Collaborations; Complex; Coupled; Data; Development; Dicer Enzyme; Disease; Enzymes; Event; Foundations; Gene Silencing; Genes; Genetic; Genetic Recombination; Genetic Screening; Goals; Health; Institution; Laboratories; Lead; Learning; Malignant Neoplasms; Massachusetts; Measures; Mediating; Messenger RNA; MicroRNAs; Microprocessor; Molecular; Nucleotides; Pathway interactions; Physiology; Plants; Play; Positioning Attribute; Post-Transcriptional Regulation; Process; Proteins; Qualifying; RNA; RNA Binding; Regulation; Reporter; Repression; Research; Ribonucleoproteins; Role; Small RNA; Structure; System; Tail; Techniques; Technology; Time; Training; Transcript; Transgenes; Translations; Universities; Untranslated RNA; Work; career; design; genome editing; genome-wide; medical schools; novel; novel therapeutic intervention; post-doctoral training; programs; protein complex; skills; temperature sensitive mutant; tenure track

 DESCRIPTION (provided by applicant): MicroRNAs are a class of endogenous small RNAs that are bound by Argonaute proteins to form the microRNA-Induced Silencing Complex (miRISC). MicroRNAs in miRISC negatively regulate complementary protein-coding genes to control many aspects of normal development. The biogenesis of microRNAs and miRISC is extensively studied, but comparatively little is understood about their turnover. Untemplated nucleotide additions at the 3' end of microRNAs are correlated with destabilization, but their functional role remains unclear. The objective of this work is to elucidate the mechanisms of microRNA and miRISC turnover, and to examine the role of 3' nucleotide additions in these processes. First, I will examine the function of candidate genes in 3' nucleotide additions and microRNA decay in the context of acute inactivation of microRNA biogenesis. Second, I will establish a system to measure the turnover of endogenously-expressed Argonaute, and use this system to assess the extent to which microRNA and miRISC turnover are coupled or independent. Third, I will conduct forward genetic screens to identify factors that impact the rate of microRNA turnover. Together, these studies will deepen our understanding of how the abundance of microRNAs and miRISC is regulated, and open up multiple new directions of research. I am uniquely qualified to conduct this research, having studied different aspects of microRNA regulation and biology throughout my graduate and postdoctoral training. In Victor Ambros's laboratory, I have used the C. elegans model system to discover a novel mechanism of post-transcriptional regulation of microRNA activity. In the proposed work, I will apply similar techniques to dissecting microRNA and miRISC turnover, while also taking advantage of the unique focus on RNA biology at the University of Massachusetts Medical School to expand my skill set through collaboration with my advisory committee. My short term career goals are to learn new techniques in biochemistry and cell biology through formal coursework and practical training, and to carry out the proposed research while transitioning to an independent position. My long term goal is to build a multi-faceted research program on the foundation of these projects in a tenure-track position at an academic institution.

 描述（由申请人提供）：MicroRNA是一类内源性小RNA，其与Argonaute蛋白结合形成microRNA诱导的沉默复合物（miRISC）。miRISC中的microRNA负调控互补蛋白编码基因，以控制正常发育的许多方面。microRNA和miRISC的生物发生被广泛研究，但对其周转率的了解相对较少。在microRNA的3'末端添加非模板核苷酸与去稳定化相关，但其功能作用仍不清楚。这项工作的目的是阐明microRNA和miRISC周转的机制，并检查3'核苷酸添加在这些过程中的作用。首先，我将研究在microRNA生物发生急性失活的背景下，候选基因在3'核苷酸添加和microRNA衰变中的功能。其次，我将建立一个系统来测量内源性表达的Argonaute的周转，并使用这个系统来评估microRNA和miRISC周转的耦合或独立程度。第三，我将进行遗传筛查，以确定影响发病率的因素。 microRNA的周转。总之，这些研究将加深我们对microRNA和miRISC丰度如何调节的理解，并开辟多个新的研究方向。我是唯一有资格进行这项研究的人，在我的研究生和博士后培训期间，我研究了microRNA调控和生物学的不同方面。在维克托·安布罗斯的实验室里，我使用了C。elegans模型系统，以发现microRNA活性的转录后调节的新机制。在拟议的工作中，我将适用类似的 技术来剖析microRNA和miRISC的周转，同时也利用马萨诸塞州医学院对RNA生物学的独特关注，通过与我的顾问委员会合作来扩展我的技能。我的短期职业目标是通过正式的课程和实践培训学习生物化学和细胞生物学的新技术，并在过渡到独立职位的同时进行拟议的研究。我的长期目标是在这些项目的基础上建立一个多方面的研究计划，在学术机构的终身职位。