Target Validation using Gene Knockouts in Somatic Cells

简介(由申请人提供)：建议进行第一阶段研究的目的 在此开发试剂和方法以快速和明确地进行筛选 将人类基因组作为药物发现目标来验证基因。在第二阶段 研究，这些方法将被用来发现和验证要追求的目标 用于治疗胰腺癌，尽管该技术适用于 其他迹象。 基因敲除一直被认为是最明确的灭活方法 基因。筛选包含失活基因的细胞库的能力 证明基因的功能已经被证明是非常成功的，但 通常是为非哺乳动物细胞保留的。创新的技术 在拟议的研究中描述的允许基因敲除和筛选 现在能够以高吞吐量执行基因评估 与哺乳动物细胞系。建议的具体目标是1)生成2,000 基因敲除基因库，2)10个基因敲除载体的创建，3) 2个人胰腺细胞系中10个基因的失活，以及4)评估 表型效应。建立大型基因库并进行筛选 定义良好的分析文库将使发现基因、他们的 生化功能及其在与人类相关的信号网络中的作用 疾病及其治疗效用。 建议的商业应用： PanGenex计划在服务能力(具有潜在的短期和长期收入预测)方面利用其创新和专有技术，并在PanGenex内部和与公司合作伙伴提供持续的药物发现计划。

DESCRIPTION (provided by applicant): The objective of Phase I studies proposed herein is to develop reagents and methods to rapidly and unambiguously screen the human genome to validate genes as drug discovery targets. In phase II studies, the methods will be used to discover and validate targets to pursue for treatment of pancreatic cancer, although the technology is applicable for other indications. Gene knockouts have been considered the most unambiguous means for inactivating genes. The ability to screen libraries of cells containing inactivated genes to demonstrate functions of genes has proven to be highly successful, but has typically been reserved for non-mammalian cells. The innovative technology described in the proposed research allows for gene knockouts and screening capabilities to enable gene evaluations to now be performed in high throughput with mammalian cell lines. Specific aims proposed are 1) generation of a 2,000 gene library for gene knockouts, 2) creation of 10 gene knockout vectors, 3) inactivation of 10 genes in 2 human pancreatic cell lines, and 4) evaluation of phenotypic effects. Creation of large libraries of genes and screening these libraries in well-defined assays will enable discovery of genes, their biochemical functions, their roles in signaling networks relevant to human diseases, and their therapeutic utilities. PROPOSED COMMERCIAL APPLICATION: PanGenex plans to capitalize on its innovative and proprietary technology both in a service capacity (with potential short-term and long-term revenue projections) and to provide for sustained drug discovery programs both within PanGenex and with corporate partners.