Structural and Mechanistic basis for RNA Silencing

RNA沉默的结构和机制基础

• 批准号: 9979882
• 负责人: IAN JOHN MACRAE
• 金额: $ 67.73万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979882
• 关键词: Biogenesis; Brain; Cell Maintenance; Cells; Complex; Development; Dicer Enzyme; Disease; Fertilization; Gene Expression; Gene Expression Regulation; Gene Silencing; Genes; Goals; Health; Homeostasis; Human; Human Biology; Human body; Investigational Therapies; Knowledge; Liquid substance; Longevity; Malignant Neoplasms; Mediating; Messenger RNA; MicroRNAs; Mutation; Names; Phase; Physiological Processes; Process; Proteins; RNA Interference; Regulation; Research; Research Personnel; Small Interfering RNA; Small RNA; Structure; Therapeutic; Work; cancer prevention; cardiogenesis; design; human disease; improved; insight; novel; programs; stem cells

Project Summary/Abstract Nearly every cell in the human body contains a set of programmable gene-silencing proteins named Argonaute. The natural function of Argonaute proteins is to mediate gene-regulation by microRNAs (miRNAs), small RNAs that contribute to cellular homeostasis during diverse physiological process, such as stem cell maintenance, fertilization, and heart development. Human Argoanute-2 (Ago2) can also be harnessed for experimental and therapeutic purposes through the introduction of small interfering RNAs (siRNAs), which are bound by Ago2 and used to direct the silencing of targeted genes. The overarching goal of this research is to understand how small RNAs are generated and used by Argonaute to silence genes. The rational motivating this work is that improved understanding of silencing processes will empower efforts to harness these mechanisms in a therapeutic setting and inform treatment of disease states resulting from aberrant miRNA function. Additionally, because miRNAs contribute in many facets of human biology, improvements in understanding function will enable researchers studying diverse aspects of human health and disease. Major projects include: 1) determining the structure of human Dicer, the enzyme that catalyzes the final step of miRNA biogenesis, with the goal of understanding how Dicer mutations drive diverse and devastating forms of human cancer; 2) identifying determinants of miRNA targeting with the goal of improving prediction of miRNA targets; 3) investigate the structure of miRNA- induced silencing complexes with a focus on understating how liquid:liquid phase separation contributes to gene-regulation by miRNAs; 4) establish the structural basis for mRNA cleavage by siRNAs, with the goal of providing novel insights for rational design of therapeutic siRNAs. The combined studies are expected to provide fundamental knowledge necessary for deciphering and controlling Argonaute- mediated regulation of human gene expression.

项目概要/摘要 人体中几乎每个细胞都含有一组可编程基因沉默蛋白，称为 阿尔戈努特。 Argonaute 蛋白的天然功能是通过 microRNA 介导基因调节 (miRNA)，在不同生理过程中有助于细胞稳态的小 RNA，例如 如干细胞维持、受精和心脏发育。人类 Argoanute-2 (Ago2) 也可以 通过引入小干扰RNA用于实验和治疗目的 (siRNA)，与 Ago2 结合并用于指导靶基因的沉默。首要的 这项研究的目标是了解 Argonaute 如何生成和使用小 RNA 来沉默 基因。这项工作的合理动机是提高对沉默过程的理解将 授权努力在治疗环境中利用这些机制并为疾病治疗提供信息 异常 miRNA 功能导致的状态。此外，由于 miRNA 在许多方面都有贡献 人类生物学，理解功能的改善将使研究人员能够研究不同的方面 人类健康和疾病。主要项目包括：1）确定人类Dicer的结构， 催化 miRNA 生物合成最后一步的酶，目的是了解 Dicer 如何 突变导致多种和毁灭性的人类癌症； 2) 鉴定miRNA的决定因素 以改善 miRNA 靶点预测为目标的靶向； 3）研究miRNA的结构- 诱导沉默复合物，重点是了解液：液相分离的贡献 通过 miRNA 进行基因调控； 4) 建立 siRNA 切割 mRNA 的结构基础，其中 目标是为治疗性 siRNA 的合理设计提供新颖的见解。合并的研究是 预计将提供破译和控制Argonaute所需的基础知识- 介导的人类基因表达调控。