Systematic Identification of Dynamic MicroRNA Targets in Development and Stress

发育和应激过程中动态 MicroRNA 靶标的系统识别

• 批准号: 7672639
• 负责人: Brian Andrew Kudlow
• 金额: $ 5.01万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7672639
• 关键词: Address; Biochemical; Biological Process; Caenorhabditis elegans; Cell physiology; Classification; Complex; Computing Methodologies; Data; Development; Diabetes Mellitus; Disease; Functional disorder; Gene Expression Regulation; Gene Silencing; Gene Targeting; Goals; Heart Diseases; Lead; Malignant Neoplasms; Maps; Mediating; Methodology; Methods; MicroRNAs; Molecular; Monitor; Process; Staging; Stress; Tissues; biological adaptation to stress; human disease; in vivo; insight; novel; response

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) regulate many biological processes, and altered miRNA expression or function is associated with human diseases including a variety of cancers, heart disease, and diabetes. However, the molecular mechanisms by which specific miRNAs regulate complex cellular and organismal functions is poorly understood, largely because very few direct targets of miRNAs have been experimentally identified. Predictions of miRNA targets by computational methods are widely variable and not yet reliable. In this proposal, I describe a novel biochemical method for the systematic and direct identification miRNA targets. Through biochemical isolation of an RNP complex, shown to be required for miRNA-mediated gene silencing, that contains both miRNAs and their actively repressed miRNA targets, I will simultaneously identify bona fide miRNA targets and monitor the dynamic expression of miRNAs. This methodology will be employed in vivo in Caenorhabditis elegans to identify miRNA targets in specific tissues, during development, and in response to organismal stresses to address the hypothesis that miRNA-mediated gene regulation is highly dynamic and tightly regulated. The studies in this proposal are intended to achieve two major goals: 1) to identify miRNAs and their targets that mediate key developmental decisions and organismal stress responses, and 2) to refine our understanding of the mechanisms that direct miRNA target gene selection. The data generated in these studies will provide insight into the mechanisms by which miRNAs regulate processes such as proliferation, differentiation, and stress responses, and how miRNA dysfunction can lead to disease states. This proposal seeks to identify mechanisms of post-transcriptional gene regulation that mediate important cellular functions such as proliferation and differentiation. Disruption of these mechanisms of gene regulation is associated with human diseases such as cancer, heart disease, and diabetes.

描述（由申请人提供）：微小RNA（miRNA）调节许多生物过程，并且改变的miRNA表达或功能与包括多种癌症、心脏病和糖尿病在内的人类疾病相关。然而，特定的miRNA调控复杂的细胞和生物功能的分子机制知之甚少，主要是因为很少有直接的靶点已经被实验确定。通过计算方法对miRNA靶点的预测变化很大，而且还不可靠。在这个建议中，我描述了一种新的生化方法，系统和直接识别miRNA的目标。通过RNP复合物的生物化学分离，显示需要miRNA介导的基因沉默，其中包含miRNA和它们的积极抑制的miRNA目标，我将同时确定真正的miRNA目标和监测的动态表达的miRNA。该方法将在秀丽隐杆线虫体内用于鉴定特定组织中的miRNA靶点，在发育过程中，以及响应生物应激，以解决miRNA介导的基因调控是高度动态和严格调控的假设。本研究旨在实现两个主要目标：1）鉴定介导关键发育决策和生物应激反应的miRNA及其靶点，2）完善我们对指导miRNA靶基因选择的机制的理解。这些研究中产生的数据将提供对miRNA调节增殖、分化和应激反应等过程的机制的深入了解，以及miRNA功能障碍如何导致疾病状态。该提案旨在确定介导重要细胞功能（如增殖和分化）的转录后基因调控机制。这些基因调控机制的破坏与人类疾病如癌症、心脏病和糖尿病有关。