Human Embryonic Stem Cell Genetic Modification Core

人类胚胎干细胞基因改造核心

• 批准号: 7356527
• 负责人: Richard R Behringer
• 金额: $ 25.37万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7356527
• 关键词: Cell Line; DNA; Doxycycline; Electroporation; Engineering; Gene Exchanges; Gene Targeting; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genotype; Goals; Growth; Human; Human Engineering; In Vitro; Individual; Karyotype; Knock-in Mouse; Knock-out; Mediating; Methods; Modification; Mus; Pharmaceutical Preparations; Pilot Projects; Plasmids; Polymerase Chain Reaction; Protein Overexpression; RNA Interference; Recombinants; Research Personnel; System; Teratoma; Tetracycline; Tetracyclines; cell bank; design; desire; embryonic stem cell; gain of function; gene induction; human embryonic stem cell; human embryonic stem cell line; in vivo; interest; programs; vector

The Genetic Modification Core will generate genetically engineered human ES (hES) cell lines required for the four primary projects and the pilot projects of the Program. These include gene null, conditional null, and knock-in hES cell lines. In addition, a very efficient recombination medicated cassette exchange method will be used to generate gene overexpression and/or knockdown (by RNAi) hES cell lines. Gene constructs generated by the Program investigators will be electroporated into NIH-approved human ES cells (H1 or H9) and cultured with appropriate drugs to select for DNA transformants. The resulting hES cell colonies will be picked and expanded for genotype analysis. Genotype analysis will be performed by the Project investigators. After the genetically engineered hES cell lines are identified and confirmed they will be transferred to Core B for karyotype analysis, differentiation potential (if required) in vitro and in vivo by teratoma formation, and expansion for cell banking. All of the four primary projects propose to generate genetically modified hES cell lines. Thus, the Genetic Modification Core is an essential component of the Program.

基因修改核心将产生基因工程人类胚胎干细胞(HES)细胞系 该方案的四个主要项目和试点项目所需的。其中包括基因缺失， 条件空的和敲入的HES细胞系。此外，一种非常有效的重组药物 将使用盒式交换法产生基因过表达和/或敲除(通过 RNAi)HES细胞株。由计划调查人员产生的基因构建将被电穿孔 进入NIH批准的人类ES细胞(H1或H9)，并与适当的药物培养以选择DNA 变形金刚。由此产生的HES细胞克隆将被挑选并扩增以进行基因分析。 基因分型将由项目调查员进行。在基因工程之后 HES细胞系被鉴定并确认将被转移到核心B进行核型分析， 通过畸胎瘤的形成和细胞的扩增在体外和体内的分化潜能(如果需要) 银行业。所有四个主要项目都提出要培育转基因的HES细胞系。因此， 基因改造核心是该计划的一个重要组成部分。