A new site specific recombinase for mammalian genome engineering

描述（由研究者提供）：位点特异性重组酶（SSR）在哺乳动物遗传分析中起关键作用。 本申请中提出的工作的中心目标是开发一种新的SSR，用于干细胞和小鼠的基因组工程。 目前，两个SSR，Cre和Flpe，被广泛使用，并已被纳入国际敲除小鼠联盟的基因打靶和基因捕获设计。 具有不同于Flpe和Cre的靶位点特异性的其他SSR将使新的策略成为可能，这是超越当前方法的重大进步。 中心目标是测试Dre重组酶作为用于ES细胞和小鼠的额外SSR的能力。 基于以前发表的工作和我实验室的新初步研究，我们知道，虽然Dre与Cre重组酶有关，但它不与细菌或哺乳动物细胞中的Cre靶位点loxP反应。 本申请中提出的工作将仔细测试Dre重组酶在干细胞和发育中的小鼠中枢神经系统中的效率和特异性。 拟议的研究将为研究界使用新的重组酶研究工具提供必要的新信息。 我们还将产生有价值的研究资源，包括在神经前体中表达Dre的小鼠品系以及Dre活性报告小鼠品系和胚胎干细胞。 公共卫生相关性拟议的实验将产生一种新的位点特异性重组酶工具，用于基因组工程应用。 这一遗传工具将有助于研究界建立人类遗传疾病和紊乱的动物模型。

DESCRIPTION (provided by investigator): Site specific recombinases (SSRs) play a critical role in mammalian genetic analysis. The central goal of the work proposed in this application is to develop a new SSR, for genome engineering in stem cells and mice. At the present time, two SSRs, Cre and Flpe, are widely used and have been incorporated into the gene targeting and gene trapping designs of the International Knockout Mouse Consortium. Additional SSRs with target site specificities differing from Flpe and Cre will enable new strategies that are a significant advancement beyond current approaches. The central goal is to test the capabilities of the Dre recombinase as an additional SSR for use in ES cells and mice. Based on previously published work and new preliminary studies in my lab, we know that although Dre is related to the Cre recombinase, it does not react with the Cre target site loxP in bacteria or in mammalian cells. The work proposed in this application will carefully test the efficiency and specificity of the Dre recombinase in stem cells and in the developing central nervous system of mice. The proposed studies will provide essential new information required for the use of a new recombinase research tool by the research community. We will also generate valuable research resources including a mouse strain that expresses Dre in neural precursors as well as a Dre activity reporter mouse strain and embryonic stem cells. PUBLIC HEALTH RELEVANCE The proposed experiments will generate a new site-specific recombinase tool for use in genome engineering applications. This genetic tool will be useful to the research community for generating animal models of human genetic diseases and disorders.