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，在其发展中发挥了重要作用的主题是，在这些途径的组成部分中描述了关键的心脏信号通路和识别，它的人类遗传多态性改变了心脏失败的易感性或进展。我们的研究者小组应用了一个系统的垂直整合分析，以识别心脏信号传导和钙处理基因中的新型多态性，从而定义了与特定的临床特征或结果的关联，从而确定了这些多态性系统的功能效应，并确定了这些多态性系统的功能效应，并确定了这些多态性系统的遗传学效应，并确定了这些遗传学系统的影响。兔子。尽管人类研究驱动了这些研究，但会清楚地提供生物学框架 机械研究，合适的小鼠模型的发展将允许实验 对人类受试者根本无法实现的单个基因变体进行。为了促进这种垂直科学方法，核心D将对小鼠基因组进行复杂的操纵，以便为SCCOR研究者的机理研究生成最佳的小鼠模型，包括条件和常规敲除，敲门式，多基因取代，多基因的取代和靶向转基因。为了支持SCCOR项目，核心将有助于设计靶向策略，将设计靶向向量，将进行电压，选择和识别靶向的ES细胞，并执行胚泡注入以生成嵌合小鼠。