Target Validation using Gene Knockouts in Somatic Cells

使用体细胞中的基因敲除进行靶标验证

• 批准号: 6484456
• 负责人: Robert E Finney
• 金额: $ 10万
• 依托单位: PANGENEX, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-08 至 2002-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6484456
• 关键词: Escherichia coli; alleles; functional /structural genomics; gel electrophoresis; gene expression; gene targeting; genetic library; genetic screening; genome; high throughput technology; molecular cloning; pancreas neoplasms; pancreatic islets; phenotype; plasmids; polymerase chain reaction; technology /technique development; tissue /cell culture; transfection

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.

描述（由申请人提供）：提议的第一阶段研究的目标 这里的目的是开发试剂和方法来快速、明确地筛选 人类基因组以验证基因作为药物发现目标。第二阶段 研究，这些方法将用于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 其他指示。 基因敲除被认为是最明确的灭活手段 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 通常是为非哺乳动物细胞保留的。创新技术 拟议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 与哺乳动物细胞系。提出的具体目标是 1) 生成 2,000 用于基因敲除的基因库，2) 创建10个基因敲除载体，3) 2 个人类胰腺细胞系中 10 个基因的失活，以及 4) 评估 表型效应。创建大型基因库并筛选这些基因 明确的分析中的文库将能够发现基因、它们的 生化功能及其在与人类相关的信号网络中的作用 疾病及其治疗用途。 拟议的商业应用： PanGenex 计划在服务能力（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 内部以及与企业合作伙伴一起提供持续的药物发现计划。