Core--Embryonic Stem Cell Gene Manipulation

在SCCOR提案的主题中，在其发展中发挥重要作用的一个主题是描述关键的心脏信号通路和鉴定，在这些通路的组成部分中，人类基因多态性改变了对心力衰竭的易感性或进展。我们的研究小组应用了系统的垂直整合分析来识别心脏信号和钙处理基因的新多态性，定义与特定临床特征或结果的关联，利用体外表达系统确定这些多态性的功能影响，并建立这些多态性在完整生物系统中的病理生理效应，即基因操纵的小鼠和兔子。尽管联合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

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.