Molecular Mechanisms of microRNA and miRISC turnover

microRNA 和 miRISC 更新的分子机制

基本信息

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
项目类别：
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&apos 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

项目摘要

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（由适用提供）：microRNA是一类由Argonaute蛋白约束以形成microRNA诱导的沉默复合物（Mirisc）的内源性小RNA。 Mirisc中的microRNA负调节完整的蛋白质编码基因，以控制正常发育的许多方面。 MicroRNA和Mirisc的生物发生是广泛研究的，但对其周转率相对较少。 MicroRNA 3'末端的未核苷酸添加与破坏稳定相关，但其功能作用尚不清楚。这项工作的目的是阐明MicroRNA和Miriscewrive的机制，并检查3'核苷酸在这些过程中的作用。首先，我将检查候选基因在3'核苷酸添加和microRNA衰减中的功能，在急性失活的microRNA生物发生情况下。其次，我将建立一个系统来衡量内源表达的Argonaute的营业额，并使用该系统来评估microRNA和Mirisce Cretplover耦合或独立的程度。第三，我将进行远期遗传筛选，以确定影响速率的因素 microRNA营业额。这些研究共同加深了我们对如何调节MicroRNA和Mirisc的丰度的理解，并打开了多个新的研究方向。在整个研究生和博士后培训中，我研究了MicroRNA调节和生物学的不同方面，因此我具有独特的资格进行这项研究。在维克多·安布罗斯（Victor Ambros）的实验室中，我使用了秀丽隐杆线虫模型系统来发现microRNA活性转录后调节的新机制。在拟议的工作中，我将应用类似的剖析MicroRNA和Mirisc流动的技术，同时还利用了马萨诸塞大学医学院对RNA生物学的独特关注，通过与我的咨询委员会的合作来扩大我的技能。我的短期职业目标是通过正式的课程和实践培训来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。我的长期目标是在学术机构的终身制位置建立一项关于这些项目基础的多面研究计划。