Core B: Gene Modulation with RNAi and CRISPER

项目总结-核心B 核心B通过提供最先进的CRISPR-Cas9和 RNAi工具。短发夹RNA（shRNAs）是在该项目的支持下开发的， 核心B和计划研究人员的创新有助于使这些极其强大的生物学 工具.在过去的资助期间，Core设计了一种预测shRNA效力的新算法， 导致了对应于注释的蛋白质编码基因的第五版RNAi文库的产生， 人类和老鼠还开发了使用CRISPR-Cas9筛选的技术，以暴露 功能重要的蛋白质结构域。在即将到来的请求支助期间，核心小组建议： 通过五个总体目标帮助项目调查人员。首先，该计划将继续提供访问状态- 现有技术的RNAi载体和CRISPR-Cas9载体以及用于分析任一单基因的方法 敲除或用于进行合并的RNAi/CRISPR筛选。第二，核心将提供方案 研究人员可以访问小鼠和人类版本5 shRNA文库，并将在 请求过程中第三，Core将构建定制的CRISPR扫描库，用于执行结构-功能分析。 分析每个程序中的感兴趣基因。第四，核心将产生定制领域为重点 CRISPR文库允许在各种情况下询问和发现癌症药物靶标。五是 Core将继续努力改进CRISPR-Cas9技术，并将这些创新技术提供给 该计划和整个科学界。

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.