Global Gene Functional Analysis with siRNA Libraries

描述(申请人提供)：由于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快药物靶点的发现和验证，并为复杂人类疾病的药物治疗和诊断开辟新的途径。最近基于小干扰RNA(SiRNA)的基因敲除技术的创新使基因功能分析发生了革命性的变化。该项目的最终目标是开发一套针对人类、小鼠和大鼠所有特有基因的小干扰RNA(SiRNA)文库，并使其在商业上可用。 该计划的第一阶段将包括开发技术，在逆转录病毒载体中构建包含8500个特征基因的高复杂性siRNA文库，并以细胞凋亡调控为模型，通过分离生物活性锥体来测试该文库的功能性能。拟议技术的一个重要技术方面是它与Affymetrix基因芯片技术的集成，这将允许使用微阵列分析的能力和速度，这种分析已经广泛地用于学术和工业研究，以发现功能基因。在第二阶段，该计划将扩展到一套人、小鼠和大鼠全基因组siRNA文库的开发和商业化。SiRNA文库将允许在全球范围内搜索与各种疾病有关的功能基因，并通过在活细胞环境中任意选择失活特定的mRNAs和相应的蛋白质来验证新的药物靶点。这项已建立的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。拟议研究的预期结果将是一套商业上可用的高通量基因功能分析试剂盒，以及用于前列腺癌治疗的有效抗癌药物靶点。

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.