MECHANISMS FOR MODULATION OF MIRNA-MEDIATED GENE SILENCING

miRNA 介导的基因沉默的调节机制

• 批准号: 9132318
• 负责人: Sergej Djuranovic
• 金额: $ 32.03万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9132318
• 关键词: 3' Untranslated Regions; Affect; Apoptosis; Base Pairing; Binding; Binding Sites; Biological; Biological Assay; Biological Process; Cell Differentiation process; Cell Proliferation; Cells; Complex; Development; Developmental Therapeutics Program; Drosophila genus; Elements; Eukaryotic Cell; Event; Fibroblasts; Gene Expression; Gene Expression Regulation; Gene Silencing; Gene Targeting; Goals; Half-Life; Health; Homeostasis; Human; In Vitro; Indium; Kinetics; Knowledge; Laboratories; Lead; Mediating; Messenger RNA; Metabolism; MicroRNAs; Molecular; Nature; Neurons; Onset of illness; Pathway interactions; Play; Post-Transcriptional Regulation; Process; Proteins; RNA; RNA-Binding Proteins; RNA-Induced Silencing Complex; Regulator Genes; Regulatory Pathway; Reporter; Reporter Genes; Repression; Research; Role; Site; System; Techniques; Time; Translation Initiation; Translational Repression; Translations; Untranslated RNA; Variant; Work; base; genome-wide; in vivo; inhibitor/antagonist; insight; interest; mRNA Decay; mRNA Stability; programs; protein complex; research study; response

 DESCRIPTION (provided by applicant): miRNAs are one of the key post-transcriptional regulators of gene expression and play a role in a wide range of biological processes including development, cell proliferation, cell differentiation, metabolism and apoptosis. We have previously deduced the mechanism and relative timing of several cellular events during miRNA- induced translational repression. Here, we will focus on elucidating mechanisms that modulate this regulatory pathway. Variations in the level of miRNA-induced gene regulation are observed in both reporter and global- based approaches, leading us to ask why and how these differences exist. Using our inducible and controllable in vivo reporter systems, as well as in vitro techniques, we will investigate the effects of mRNA target intrinsic stability on the kinetic and dissect contributions of miRICS components in the process of miRNA-mediated gene silencing. We will employ this system with a variety of modified and natural reporter genes to get a more complete picture of the relationships between distinct RNA binding proteins and miRNA pathway components on gene regulation. We will use our newly developed assays and techniques to deduce the molecular mechanisms for modulation of miRNA-mediated gene silencing by these RNA-binding proteins. Finally, these studies will be followed by genome-wide approaches in a biological context that will allow us to investigate the importance of cis and trans factors on the modulation of specific miRNA response across endogenous gene targets. In addition to vastly expanding our knowledge about miRNA-mediated gene silencing, the proposed work will give us more clues as to how alterations in systems that modulate miRNA response may influence cellular homeostasis in regular and pathological states.

 描述（由申请人提供）：miRNA是基因表达的关键转录后调节因子之一，并在包括发育、细胞增殖、细胞分化、代谢和凋亡在内的广泛的生物过程中发挥作用。我们先前已经推断了在miRNA诱导的翻译抑制过程中几个细胞事件的机制和相对时间。在这里，我们将重点阐明调节这一调节途径的机制。在基于报告基因和基于全局的方法中观察到miRNA诱导的基因调控水平的变化，这使我们询问为什么以及如何存在这些差异。使用我们的可诱导和可控的体内报告系统，以及在体外技术，我们将研究mRNA靶的内在稳定性的动力学和解剖贡献的miRICS组件在miRNA介导的基因沉默的过程中的影响。我们将使用这个系统与各种修饰和天然的报告基因，以获得不同的RNA结合蛋白和miRNA通路的基因调控组件之间的关系更完整的图片。我们将使用我们新开发的检测方法和技术来推断这些RNA结合蛋白调节miRNA介导的基因沉默的分子机制。最后，这些研究将遵循生物学背景下的全基因组方法，这将使我们能够研究顺式和反式因子对内源性基因靶标的特异性miRNA应答的调节的重要性。除了极大地扩展我们对miRNA介导的基因沉默的了解之外，这项拟议的工作还将为我们提供更多线索，以了解调节miRNA反应的系统的改变如何影响正常和病理状态下的细胞稳态。