Core--Embryonic Stem Cell Gene Manipulation

核心--胚胎干细胞基因操控

• 批准号: 7312581
• 负责人: Gerald W. Dorn
• 金额: $ 43.29万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7312581
• 关键词: artificial chromosomes; biomedical facility; biotechnology; disease /disorder model; electroporation; embryonic stem cell; experimental designs; gene expression; gene targeting; genetic manipulation; genetically modified animals; heart failure; laboratory mouse; model design /development; transfection /expression vector

Among the themes of this SCCOR proposal, one that has played a major role in its development is the delineation of critical cardiac signaling pathways and identification, in the components of these pathways, of human genetic polymorphisms that modify the susceptibility to, or progression of, heart failure. Our group of investigators has applied a systematic vertically-integrated analysis for identifying novel polymorphisms in cardiac signaling and calcium-handling genes, defining associations with specific clinical features or outcomes, determining the functional effects of these polymorphisms using in vitro expression systems, and establishing the pathophysiological effects of these polymorphisms in intact biological systems, i.e. genetically manipulated mice and rabbits. Although associative and case-control human studies drive these investigations, and will clearly provide the biological framework for mechanistic studies, the development of suitable mouse models will permit experiments to be performed on individual gene variants that are simply not attainable with human subjects. To facilitate this vertical scientific approach, Core D will perform complex manipulations of the mouse genome in order to generate optimal mouse models for the SCCOR investigators' mechanistic studies, including conditional and conventional knock-outs, knock-ins, multi-gene substitutions, and targeted transgenesis. In support of the SCCOR projects, the Core will assist in the design of targeting strategies, will engineer targeting vectors, will electroporate, select for and identify targeted ES cells, and will perform blastocyst injection for generation of chimeric mice.

在该 SCCOR 提案的主题中，在其发展中发挥重要作用的一个主题是界定关键的心脏信号传导途径，并在这些途径的组成部分中识别人类遗传多态性，这些多态性会改变心力衰竭的易感性或进展。我们的研究小组应用系统的垂直整合分析来识别心脏信号和钙处理基因中的新多态性，定义与特定临床特征或结果的关联，使用体外表达系统确定这些多态性的功能效应，并确定这些多态性在完整生物系统（即基因操纵的小鼠和小鼠）中的病理生理学效应。 兔子。尽管联想和病例对照人类研究推动了这些调查，并将明确提供生物学框架 机制研究，合适的小鼠模型的开发将使实验得以进行 对人类受试者根本无法实现的个体基因变异进行了研究。为了促进这种垂直科学方法，Core D将对小鼠基因组进行复杂的操作，以便为SCCOR研究人员的机制研究生成最佳小鼠模型，包括条件和常规敲除、敲入、多基因替换和靶向转基因。为了支持 SCCOR 项目，核心将协助设计靶向策略、设计靶向载体、电穿孔、选择和识别靶向 ES 细胞，并进行囊胚注射以产生嵌合小鼠。