Modulation of Gene Expression Through RNAi

核心B通过提供对最先进的RNAi工具的访问来支持该计划内的每个项目。短的 发夹RNA(ShRNA)是在该计划的支持下开发的，Core正在进行创新 B和项目调查人员帮助制造了这些极其强大的生物工具。在过去 在资助期间，Core设计了快速设计携带受调控shRNA表达的小鼠的方法 盒式磁带，设计了以多路复用方式在功能上验证shRNA的策略，并创建了第三个 在人类和小鼠中生成与注释的蛋白质编码基因相对应的RNAi文库。在.期间 在即将到来的请求支持期间，核心建议通过五个阶段来帮助方案调查人员 总体目标。首先，该计划将继续提供对最先进的RNAi工具的访问，以进行分析 要么是单基因敲除，要么是进行RNAi筛选。第二，核心将与 研究人员操作“传感器分析”以确定针对感兴趣基因的最佳shRNAs工具。第三, 核心将根据程序感兴趣的基因集产生定制的RNAi文库 调查人员。第四，核心将生产携带受监管的RNAi盒式磁带的小鼠，并帮助调查人员 将这些与其他所需的基因损伤结合起来。最后，核心将继续努力改进RNAi 技术，并将这些创新提供给该计划和整个社区。

Core B supports every project within the Program by providing access to state-of-the-art 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 methods to rapidly engineer mice carrying regulated shRNA expression cassettes, devised strategies for functionally validating shRNAs in a multiplexed fashion, and created a third generation of RNAi libraries corresponding to annotated protein coding genes in humans and mice. During the upcoming period of requested support, the core proposes to aid Program investigators through five general aims. First, the program will continue to provide access to state-of-the-art RNAi tools for analyzing either single gene knockdowns or for performing pooled RNAi screens. Second, the Core will work with investigators to operate the "sensor assay" to identify optimal shRNAs tools against genes of interest. Third, the core will produce custom RNAi libraries against sets of genes that are of interest to Program investigators. Fourth, the Core will produce mice carrying regulated RNAi cassettes and aid investigators in combining these with other desired genetic lesions. Finally, the Core will carry on its efforts to improve RNAi technologies and make those innovations available to the Program and to the community at large.