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）：提出的第一阶段研究的目的 本文是开发试剂和方法来快速，明确筛选 人类基因组验证基因作为药物发现靶标。在第二阶段 研究，这些方法将用于发现和验证目标以追求目标 用于治疗胰腺癌，尽管该技术适用于 其他迹象。 基因敲除被认为是灭活的最明确手段 基因。筛选包含灭活基因的细胞库的能力 证明基因的功能已被证明是非常成功的，但是 通常保留给非哺乳动物细胞。创新技术 拟议的研究中描述的允许进行基因敲除和筛查 现在可以在高通量中进行基因评估的能力 与哺乳动物细胞系。提出的具体目标是1）生成2,000 基因敲除基因库，2）创建10个基因敲除载体，3） 在2种人胰腺细胞系中灭活10个基因，4）评估 表型效应。创建大型基因图书馆并筛选这些 定义明确的测定中的图书馆将发现基因 生化功能，它们在与人类相关的信号网络中的作用 疾病及其治疗性公用事业。 拟议的商业应用： Pangenex计划以服务能力（潜在的短期和长期收入预测）来利用其创新和专有技术，并为Pangenex和公司合作伙伴提供持续的药物发现计划。