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用于操作小鼠基因组的位点特异性重组酶。

Site-specific recombinases for manipulation of the mouse genome.

作者信息

Birling Marie-Christine, Gofflot Françoise, Warot Xavier

机构信息

Institut Clinique de la Souris - Mouse Clinical Institute (ICS-MCI), Illkirch Cedex, France.

出版信息

Methods Mol Biol. 2009;561:245-63. doi: 10.1007/978-1-60327-019-9_16.

Abstract

Site-specific recombination systems are widespread and popular tools for all scientists interested in manipulating the mouse genome. In this chapter, we focus on the use of site-specific recombinases (SSR) to unravel the function of genes of the mouse. In the first part, we review the most commonly used SSR, Cre and Flp, as well as the newly developed systems such as Dre and PhiC31, and we present the inducible SSR systems. As experience has shown that these systems are not as straightforward as expected, particular attention is paid to facts and artefacts associated with their production and applications to study the mouse genome. In the next part of this chapter, we illustrate new applications of SSRs that allow engineering of the mouse genome with more and more precision, including the FLEX and the RMCE strategies. We conclude and suggest a workflow procedure that can be followed when using SSR to create your mouse model of interest. Together, these strategies and procedures provide the basis for a wide variety of studies that will ultimately lead to the analysis of the function of a gene at the cellular level in the mouse.

摘要

位点特异性重组系统是所有对操纵小鼠基因组感兴趣的科学家广泛使用且十分受欢迎的工具。在本章中,我们重点关注使用位点特异性重组酶(SSR)来阐明小鼠基因的功能。在第一部分,我们回顾了最常用的SSR,即Cre和Flp,以及新开发的系统,如Dre和PhiC31,并介绍了可诱导的SSR系统。经验表明,这些系统并不像预期的那么简单,因此我们特别关注与它们在小鼠基因组研究中的产生和应用相关的事实和假象。在本章的下一部分,我们阐述了SSR的新应用,这些应用能够越来越精确地对小鼠基因组进行工程改造,包括FLEX和RMCE策略。我们进行总结并提出一个工作流程,在使用SSR创建感兴趣的小鼠模型时可以遵循该流程。总之,这些策略和程序为各种各样的研究提供了基础,最终将促成在小鼠细胞水平上对基因功能的分析。

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