CCR RNAi initiative overview

基因沉默组（GSS）旨在成为CCR内研究和应用RNAi在哺乳动物细胞中的卓越和专业知识的中心。RNAi领域的科学和临床发展正在迅速影响我们对RNA沉默机制在控制正常和疾病相关基因表达（包括癌症）中的作用的理解。此外，利用RNAi分析研究基因功能的技术发展、新模型系统的开发、对药物：靶标相互作用的理解的提高以及通过大规模筛选（包括全基因组）进行靶标鉴定，RNAi方法（siRNA和shRNA）现在是非常现实的。GSS继续以多种方式为CCR RNAi倡议做出贡献，包括：(a)伸出援助之手：我们已经向感兴趣的CCR实验室、分支机构和CCR学院/工作组/项目小组提供了许多讲座，目的是教育个人了解RNAi领域的现状和GSS的具体作用。曾为多个CCR实验室和分支机构的不同程度的研究人员提供咨询，包括CTB、CCBB、EIB、GB、HAMB、NOB、POB、大华银行、LBC、LCP、LCB、LCRC、LCMB、LCCTP、LCO、LEC、LEI、LG、LHC、LICB、LM、LMB、LMI、LTIB、LMP、LP、LPG、SBL。(B)信息技术：以计算机为基础获取与rna重组有关的信息将是传播rna重组技术的关键。我们现在定期更新到GSS网站，在OSTP， OD， CCR网站上报告siRNA表征数据，这些数据可供CCR研究人员使用，我们设想不久将引入描述shRNA资源和标准化协议访问的页面。此外，capplen博士还参与了NCBI中央RNAi数据库的开发，该数据库将于2005年秋季全面投入使用。(C)初始基因靶点和扩展到全基因组分析：在2003年秋季和2004年初，开展了一项广泛的外展计划，以确定关键基因靶点，其中RNAi分析将提高我们对基因/蛋白质在癌症过程中或作为抗癌靶点的作用的理解。正如下文和其他项目中所描述的那样，GSS现在已经建立了RNAi资源，适当的分析和下游功能分析，用于这些目标的很大一部分。这一初步分析的成功现在给了我们足够的信心，开始扩大我们的职权范围，考虑全基因组RNAi方法。(D) RNAi资源：GSS目前可获得约400种癌症相关基因的合成sirna，我们已经完成了85种这些基因对应的sirna介导的RNAi的表征。CCR研究人员可以使用内部MTA机制访问这些表征数据。在2005财年，我们购买了300个shRNA克隆来补充我们的合成siRNA资源，以支持一个特定的合作项目。最近在校外社区开发和可购买的shRNA资源方面取得的进展使我们有足够的信心投资于全基因组人类shRNA文库。该图书馆作为一个工作实体的建立工作将于2005年秋季开始。(E)与其他CCR计划的关系：我们也继续与其他CCR计划、计划、院系和团体进行互动，包括MTDP/MTP、DTP、RTP、MWG、ABCC和AMI。

The Gene Silencing Section (GSS) aims to be a center of excellence and expertise within CCR for the study and application of RNAi in mammalian cells. The scientific and clinical developments in the field of RNAi are rapidly impacting our understanding of the role of RNA silencing mechanisms on the control of normal and disease related gene expression, including cancer. Further, technological developments utilizing RNAi analysis to study gene function, the development of new model systems, improved understanding of drug: target interactions and target identification through large scale-screening, including whole genome, RNAi approaches (siRNA and shRNA) are now highly realistic. The GSS continues to contribute the CCR RNAi initiative in a number of ways including: (A) Out reach and assistance: We have delivered a number of lectures to interested CCR laboratories, Branches and CCR Faculty/Working Group/Program based groups with the aim of educating individuals as to the current status of the RNAi field and the role of GSS specifically. Time has been spent advising to various degrees investigators from many CCR laboratories and Branches including, CTB, CCBB, EIB, GB, HAMB, NOB, POB, UOB, LBC, LCP, LCB, LCRC, LCMB, LCCTP, LCO, LEC, LEI, LG, LHC, LICB, LM, LMB, LMI, LTIB, LMP, LP, LPG, SBL. (B) Information technology: Critical to the dissemination of RNAi technology will be computer-based access to RNAi-related information. We our now posting regular updates to the GSS website within the OSTP, OD, CCR web-site reporting siRNA characterization data that is available to CCR investigators and we envisage shortly introducing pages describing access to shRNA resources and standardized protocols. In addition, Dr. Caplen has had input into the development of a centralized RNAi database at NCBI which should become fully operational in Fall 2005. (C) Initial Gene Targets and expansion to whole genome analysis: During Fall 2003 and early 2004 an extensive out-reach program was conducted to identify key gene targets where RNAi analysis would improve our understanding of the role of a gene/protein in the cancer process and or as an anti-cancer target. As will be described below and within other projects GSS has now established RNAi resources, appropriate assays and down-stream functional analysis for a significant proportion of these targets. Success with this initial analysis has now given us sufficient confidence to begin the process of expanding our remit to consider whole genome RNAi approaches. (D) RNAi resources: Synthetic siRNAs to approximately 400 cancer-associated genes are now available to GSS and we have completed characterization of the RNAi mediated by the siRNAs corresponding to 85 of these genes. CCR investigators can access this characterization data using an intramural MTA mechanism. During FY05 we supplemented our synthetic siRNA resources with 300 shRNA clones purchased to support a specific collaborative project. Recent advancements in shRNA resources developed by the extramural community and available through purchase have given us sufficient confidence to invest in a whole genome human shRNA library. Work on the establishment of this library as a working entity will begin in Fall 2005. (E) Relationship to other CCR initiatives: We continue to also interact with other CCR initiatives, programs, faculties and groups including MTDP/MTP, DTP, RTP, MWG, ABCC and the AMI.