Global Gene Functional Analysis with siRNA Libraries

使用 siRNA 文库进行全局基因功能分析

• 批准号: 6736681
• 负责人: ALEX CHENCHIK
• 金额: $ 14.74万
• 依托单位: SYSTEM BIOSCIENCES, LLC (SBI)
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2004-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6736681
• 关键词: CD antigens; RNA interference; animal genetic material tag; expression cloning; functional /structural genomics; gene expression; gene expression profiling; genetic library; human genetic material tag; microarray technology; small interfering RNA; technology /technique development; transfection; transfection /expression vector; tumor necrosis factor alpha

DESCRIPTION (provided by applicant): Owing to the massive amount of data accumulated through various genome-wide sequencing and gene expression projects, there is an urgent need to develop novel tools to uncover the functions of each gene in human genome. Functional genomics studies will speed up the discovery and validation of drug targets, and open avenues for new concepts in drug therapy and diagnostics of complex human diseases. Recent innovations in gene knockdown technology based on small interfering RNA (siRNA) have revolutionized gene functional analysis. The ultimate goal of the proposed project is to develop and make commercially available a set of libraries expressing small interfering RNA (siRNA) against all characterized human, mouse and rat genes. Phase I of this program will cover development of technology for construction of high complexity siRNA library of 8,500 characterized human genes in retroviral vector and testing functional performance of this library by isolation of biologically active cones using apoptosis modulation as a model. An important technical aspect of the proposed technology is its integration with the Affymetrix GeneChip technology that would allow to use the power and speed of microarray analysis that is already broadly available for academic and industrial research for functional gene discovery purposes. At the Phase II, the program will be extended towards development and commercialization of a set of human, mouse and rat whole-genome siRNA libraries. siRNA libraries will allow to perform global search for functional genes involved in a variety of diseases and to validate new drug targets by arbitrary selective inactivation of specific mRNAs and corresponding proteins in the context of the living cells. The established technology will be applied, in collaboration with Cleveland Clinic Foundation, to drug target discovery in prostate cancer. The anticipated outcomes of the proposed research will be a commercially available set of kits for high-throughput gene functional analysis and validated anticancer drug targets for treatment of prostate cancer.

描述（申请人提供）：由于通过各种全基因组测序和基因表达项目积累了大量数据，迫切需要开发新的工具来揭示人类基因组中每个基因的功能。功能基因组学研究将加速药物靶点的发现和验证，并为药物治疗和诊断复杂人类疾病的新概念开辟道路。基于小干扰RNA（siRNA）的基因敲除技术的最新创新已经彻底改变了基因功能分析。拟议项目的最终目标是开发并商业化一套表达针对所有人类、小鼠和大鼠特征基因的小干扰RNA（siRNA）的文库。 该计划的第一阶段将包括在逆转录病毒载体中构建8，500个特征化人类基因的高复杂性siRNA文库的技术开发，并通过使用细胞凋亡调节作为模型分离生物活性锥来测试该文库的功能性能。所提出的技术的一个重要的技术方面是其与Affyssin基因芯片技术的集成，该技术将允许使用微阵列分析的能力和速度，该微阵列分析已经广泛用于学术和工业研究以用于功能基因发现目的。在第二阶段，该计划将扩展到一套人类，小鼠和大鼠全基因组siRNA文库的开发和商业化。siRNA文库将允许进行涉及各种疾病的功能基因的全局搜索，并通过在活细胞的背景下任意选择性地失活特定mRNA和相应蛋白质来验证新的药物靶标。该技术将与克利夫兰临床基金会合作应用于前列腺癌的药物靶点发现。拟议研究的预期成果将是一套用于高通量基因功能分析和经验证的前列腺癌治疗抗癌药物靶点的市售试剂盒。