Mechanistic Investigation of RNA-Mediated Gene Regulation and Immunity

项目摘要 拟议中的研究结合了生物物理学、分子遗传学和基因组学工具， RNA主导的生物过程的机制。我们的研究集中在RNA生物学的两个中心主题。 (1)RNA可以作为信息载体引导蛋白质的作用。这样的系统本质上是精确的 和可编程的，因此在生物技术和医学方面具有巨大的潜力，如RNAi和 CRISPR-Cas.我的实验室花了十多年的时间来剖析CRISPR-Cas的机制， 一直在基于我们的机械见解开发基因组编辑应用程序。(2)RNA可以通过定义 三级结构发挥重要作用。我的实验室已经阐明了 许多核糖开关的动力学。高分辨率的信息可能有助于开发新的抗生素。我们 现在，我们的目标是将联合收割机遗传筛选与生化纯化相结合，以系统地鉴定孤儿配体 核糖开关这些研究经常揭示细菌生理学的新方面，这些方面甚至可能是保守的， 真核生物我们建议在以下四个方面开展研究：1）结构，机制和基因组 I型CRISPR（CRISPR-Cas 3）的编辑应用; 2）CRISPR免疫建立的机制， Cas 1-Cas 2整合酶; 3）CRISPR-Tn 7的机制和基因组编辑应用; 4）解剖核糖开关- 控制机制和细菌生理学。

Project Summary The proposed research combines biophysics, molecular genetics, and genomics tools to provide high-resolution mechanisms for RNA-dominated biological processes. Our study focuses on two central themes in RNA biology. (1) RNA can serve as an information carrier to guide the action of proteins. Such systems are inherently precise and programmable, therefore carry great potential in biotechnology and medicine, as exemplified by RNAi and CRISPR-Cas. My lab spent more than ten years dissecting the mechanisms of CRISPR-Cas, and more recently has been developing genome editing applications based on our mechanistic insights. (2) RNA can adopt defined tertiary structures to exert important functions. My lab has elucidated the structure, function, and conformation dynamics of many riboswitches. The high-resolution information may enable new antibiotics development. We now aim to combine genetic screen with biochemical purification to systematically identify ligands for orphan riboswitches. Such studies often reveal novel aspects of bacterial physiology, which may even be conserved in eukaryotes. We propose to work on the following four areas of research: 1) Structure, mechanism, and genome editing applications of Type I CRISPR (CRISPR-Cas3); 2) Mechanism of CRISPR immunity establishment by Cas1-Cas2 integrase; 3) Mechanism and genome editing application of CRISPR-Tn7; 4) Dissecting riboswitch- controlled mechanism and bacterial physiology.