Core B: Gene Modulation with RNAi and CRISPER

项目摘要--核心B 核心B通过提供对最先进的CRISPR-CAS9和 RNAi工具。短发夹状RNA(ShRNAs)是在该计划的支持下开发的，并正在进行中 Core B和项目调查人员的创新帮助使这些极其强大的生物 工具。在过去的资助期间，Core设计了一种新的算法来预测shRNA的效力，该算法 导致了与注释的蛋白质编码基因相对应的第5版RNAi文库的产生 人类和老鼠。还开发了使用CRISPR-CAS9筛查来曝光的技术 具有重要功能的蛋白质结构域。在即将到来的请求支持期间，核心计划提议 援助计划的调查人员通过五个总体目标。首先，该计划将继续提供访问州- 最新的RNAi载体和CRISPR-Cas9载体以及分析任一单基因的程序 击倒或用于执行池RNAi/CRISPR筛选。第二，核心将提供计划 能够访问小鼠和人类版本5 shRNA文库的研究人员，并将在 请求。第三，核心将构建用于执行结构功能的定制CRISPR扫描库 对每个项目中感兴趣的基因进行分析。第四，核心将产生以领域为重点的定制 CRISPR图书馆，允许在各种情况下审问和发现抗癌药物靶标。第五， CORE将继续努力改进CRISPR-CAS9技术，并将这些创新提供给 该计划并向整个科学界致敬。

PROJECT SUMMARY-CORE B Core B supports every project within the Program by providing access to state-of-the-art CRISPR-Cas9 and RNAi tools. Short hairpin RNAs (shRNAs) were developed with the support of this program, and ongoing innovations by Core B and Program investigators have helped to make these extremely powerful biological tools. During the past funding period, the Core devised a novel algorithm for predicting shRNA potency, which led to the generation of a 5th version of RNAi libraries corresponding to annotated protein coding genes in humans and mice. Technology has also been developed for using CRISPR-Cas9 screening to expose functionally important protein domains. During the upcoming period of requested support, the Core proposes to aid Program investigators through five general aims. First, the program will continue providing access to state- of-the-art RNAi vectors and CRISPR-Cas9 vectors and procedures for analyzing either single gene knockdowns or for performing pooled RNAi/CRISPR screens. Second, the Core will provide Program investigators with access to the mouse and human version 5 shRNA library and will compile sub-libraries upon request. Third, the Core will construct custom CRISPR-scanning libraries for performing structure-function analysis on genes of interest within each Program. Fourth, the Core will produce custom domain-focused CRISPR libraries to allow for interrogation and discovery cancer drug targets in various contexts. Fifth, the Core will carry on its efforts to improve CRISPR-Cas9 technologies, and make those innovations available to the Program and to the scientific community at large.