Trans-NIH RNAi Facility (TNRF)

小干扰RNA(SiRNA)分子是一段RNA，通过一种称为RNA干扰(RNAi)的自然过程来阻止基因的活动。RNAi已经成为世界各地数以千计的实验室用来了解基因功能的强大工具。通过阻断或调节基因功能，RNAi可以告诉我们任何基因在维持健康或导致疾病方面的作用，这是识别潜在药物靶点的宝贵一步。在被称为全基因组RNAi筛选的测试中，科学家使用自动化将siRNAs引入人类细胞，以阻止每个基因的活动，一次一个。这一过程可以产生一份涉及特定生物功能或疾病过程的所有基因的完整列表。科学家还可以使用这些技术来了解基因在药物有效性中扮演的角色。 由于缺乏进行全基因组RNAi筛选的专业知识，缺乏能够正确解释这些实验的方法，以及公共数据库中缺乏可供研究人员参考的全面RNAi数据，RNAIS的潜在有用性一直受到限制。为了解决这些问题，NCATS运营着最先进的RNAi筛查设施，NCATS工作人员协助NIH内部调查人员进行项目规划和执行的所有阶段。该倡议通过国家医学图书馆PubChem数据库向公众提供从这些实验中产生的RNAi数据。此外，私营部门的生物技术合作伙伴也提供了siRNA序列信息。例如，研究人员可以访问Life Technologies Silencer Select siRNA文库，其中包括针对20,000多个人类基因的65,000个siRNA序列。RNAi高通量筛查设施由NCATS临床前创新人员事业部管理，提供一个带有集成自动化设备的机器人平台，用于执行筛查分析(测试)的所有方面，包括操纵化学品和细胞、读取结果并对细胞进行成像。离线(非机器人)设备可以执行从化验优化到中等规模筛选的较小规模的工作。研究人员可以选择使用几种不同的siRNA文库和其他参与RNAi的小分子。对于数据分析，该设施提供了强大的计算工具。 除了在特定项目上实现合作外，RNAi设施的工作人员还致力于开发方法，以推进RNAi筛选和信息学的科学，并追求探索基因功能的新技术。目前，NCATS的科学家们正在致力于开发一个平台，提供补充的高通量基因组技术，以推进功能基因组学领域。其目标是加快在所有疾病领域的科学发现。项目领域包括癌症(药物增强剂/耐药性筛查、三维转移筛查、癌症的分子靶点和癌症相关途径)、传染病(病毒感染和复制，如艾滋病毒、埃博拉病毒和丙型肝炎病毒)、基础细胞生物学(DNA复制和重新编程/分化)以及其他与疾病相关的表型(帕金森氏病、糖尿病和脆性X综合征)。

Small interfering RNA (siRNA) molecules are pieces of RNA that block the activity of genes through a natural process called RNA interference (RNAi). RNAi has emerged as a powerful tool used in thousands of labs worldwide to understand gene function. By blocking or modulating a genes function, RNAi can tell us about the role of any gene in maintaining health or causing disease, an invaluable step in identifying potential drug targets. In tests called genome-wide RNAi screens, scientists use automation to introduce siRNAs into human cells to block the activity of each gene, one at a time. This process can produce a complete list of all genes involved in a particular biological function or disease process. Scientists also can use these techniques to understand what roles genes play in drug effectiveness. RNAis potential usefulness has been limited by the lack of expertise to perform genome-wide RNAi screens, the lack of methodologies that can properly interpret these experiments and the absence of comprehensive RNAi data in public databases for researchers to reference. To address these problems, NCATS operates a state-of-the-art RNAi screening facility, and NCATS staff assists NIH intramural investigators with all stages of project planning and execution. The initiative provides public access to RNAi data generated from these experiments through the National Library of Medicines PubChem database. In addition, siRNA sequence information is available from private-sector biotechnology partners. For instance, researchers can access Life Technologies Silencer Select siRNA library, which includes 65,000 siRNA sequences that target more than 20,000 human genes. The RNAi high-throughput screening facility, administered by NCATS Division of Pre-Clinical Innovation staff, offers a robotic platform with integrated, automated devices for conducting all aspects of screening assays (tests), including manipulating chemicals and cells, reading the results and imaging the cells. Offline (non-robotic) devices can perform smaller-scale work from assay optimization through medium-scale screening. Investigators have the option of using several different siRNA libraries and other small molecules involved in RNAi. For data analysis, the facility offers powerful computational tools. In addition to enabling collaborations on specific projects, RNAi facility staff work to develop methods that advance the science of RNAi screening and informatics and pursue new technologies for exploring gene function. Currently, NCATS scientists are working on developing a platform that offers complementary high-throughput genomic technologies to advance the field of functional genomics. The goal is to speed scientific discoveries in all disease areas. Project areas include cancer (drug enhancer/resistance screens, development of 3-D metastasis screens, molecular targets in cancer and cancer-related pathways), infectious diseases (viral infection and replication such as HIV, Ebola virus and Hepatitis C virus), fundamental cell biology (DNA replication and reprogramming/differentiation), and other disease-related phenotypes (Parkinsons disease, diabetes and fragile X syndrome).