Integrated instrument system for maintenance and delivery of RNAi libraries

描述(申请人提供)：RNA干扰作为哺乳动物细胞中转录后基因功能沉默的一种机制的出现，代表着基因功能实验分析的巨大进步和巨大机遇。随着siRNAs和shRNAs全基因组文库的发展，我们已经进入了一个新的时代，在这个时代，强大的正向遗传学方法的应用可以在体内和体外为基因分配功能，描绘这些基因影响正常和疾病细胞过程的分子途径，并为开发新的和改进现有的治疗方法提供必要的知识。杜克大学RNA生物学中心、杜克综合癌症中心和杜克大学基因组科学与政策研究所联合起来，建立了一个RNAi共享资源设施。RNAi设施的主要任务是开发和部署RNAi技术，以支持杜克大学的功能基因组研究计划。通过对RNAi和互补技术的大量投资，我们正在为功能基因组学组装最先进的基础设施，为研究人员提供访问全基因组RNAi试剂和进行大规模功能丧失研究所需的基础设施。在这项提案中，我们寻求支持，通过收购用于RNAi/小分子文库的维护和交付的Velocity 11集成仪器系统来显著增强这一技术基础设施。该仪器系统将提供RNAi文库格式化、平板处理、液体分配、平板密封和平板识别/管理所需的自动化。该仪器将完成一套必要的工具，使基于全基因组RNAi的筛查成为杜克大学研究界的现实。Velocity 11系统从一开始就被设计为一种完全集成的计算机控制仪器，并由Velocity 11预先配置，用于通过哺乳动物细胞的脂质基转染或病毒转导来维护和传递siRNA文库所需的多种工作流程选项，同时保持在小分子文库筛选中有用的灵活性。与我们现有的下游基于高含量细胞的成像系统相结合，基于全基因组RNAi的筛选代表着一项巨大的技术进步，使研究人员能够识别在几乎任何正常或疾病相关细胞过程中发挥作用的基因。

DESCRIPTION (provided by applicant): The advent of RNA interference as a mechanism for post-transcriptional silencing of gene function in mammalian cells represents a quantum advance and tremendous opportunity for experimental analysis of gene function. With the development of genome-wide libraries of siRNAs and shRNAs, we have entered a new era in which the application of powerful forward genetic approaches can be used both in vivo and in vitro to assign function to genes, delineate molecular pathways in which these genes affect normal and disease cellular processes, and contribute to the knowledge necessary to develop new and improve existing therapies. The Duke Center for RNA Biology, the Duke Comprehensive Cancer Center, and the Duke Institute for Genome Sciences and Policy, have come together to establish an RNAi shared-resource facility. A primary mission of the RNAi Facility is to develop and deploy RNAi technologies to support functional genomics research programs at Duke. Through large investments in RNAi and complementary technologies, we are assembling a state-of-the-art infrastructure for functional genomics, providing researchers access to genome- wide RNAi reagents and the infrastructure necessary to conduct large-scale loss-of-function studies. In this proposal, we seek support to significantly enhance this technology infrastructure with the acquisition of the Velocity 11 Integrated Instrument System for Maintenance and Delivery of RNAi/Small Molecule Libraries. This instrument system will provide the automation required for RNAi library formatting, plate handling, liquid dispensing, plate sealing and plate identification/management. This instrument will complete the suite of tools necessary to make whole genome RNAi based screening a reality for the Duke research community. The Velocity 11 system was designed from the ground up as a fully integrated computer controlled instrument and is pre-configured by Velocity 11 for the multiple workflow options necessary to maintain and deliver siRNA libraries via lipid based transfection or viral based transduction of mammalian cells while maintaining the flexibility to be useful in small molecule library screening. Coupled with our existing downstream High-content cell based imaging system, whole genome RNAi based screening represents a tremendous technological advance, enabling researchers to identify genes with roles in virtually any normal or disease-associated cellular process.