Target Validation using Gene Knockouts in Somatic Cells

描述（由申请人提供）：拟定的I期研究目的 本发明目的是开发试剂和方法以快速和明确地筛选 人类基因组来验证基因作为药物发现靶点。在第二阶段 研究，这些方法将用于发现和验证目标追求 用于治疗胰腺癌，尽管该技术适用于 其他迹象。 基因敲除已被认为是失活的最明确手段 基因.筛选含有失活基因的细胞文库的能力， 证明基因的功能已被证明是非常成功的，但 通常只用于非哺乳动物细胞。创新技术 在拟议的研究中描述的允许基因敲除和筛选 现在能够以高通量进行基因评估的能力 哺乳动物细胞系。提出的具体目标是：1）2000年的一代 用于基因敲除的基因文库，2）创建10个基因敲除载体，3） 在2个人胰腺细胞系中10个基因的失活，和4）评估 表型效应创建大型基因文库并筛选这些 明确定义的测定中的文库将能够发现基因， 生物化学功能，它们在与人类相关的信号网络中的作用 疾病及其治疗效用。 拟定商业应用： PanGenex计划利用其创新和专有技术提供服务（具有潜在的短期和长期收入预测），并在PanGenex内部和与企业合作伙伴一起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.