cre - loxp介导的重组:一般原理和实验考虑

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2017-03-02 DOI:10.1002/cpmo.22

Micheal A. McLellan, Nadia A. Rosenthal, Alexander R. Pinto

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引用次数: 110

摘要

cre-loxP介导的重组系统(“cre-loxP系统”)是哺乳动物遗传学和细胞生物学不可或缺的实验工具。该系统的使用极大地扩展了我们精确询问小鼠基因功能的能力，提供了基因表达的空间和时间控制。这在很大程度上是由于其使用的简单性和对解决各种生物学问题的适应性。虽然cre-loxP系统的使用越来越广泛，特别是因为条件小鼠突变体的可用性越来越高，但在使用cre-loxP系统时需要考虑许多因素。本文综述了cre-loxP系统及其各种排列。它解决了cre-loxP技术的局限性和实验设计的相关考虑，并讨论了位点特异性基因重组和整合的替代策略。©2017 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Cre-loxP-Mediated Recombination: General Principles and Experimental Considerations

The cre-loxP–mediated recombination system (the “cre-loxP system”) is an integral experimental tool for mammalian genetics and cell biology. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. This has been largely due to the simplicity of its use and its adaptability to address diverse biological questions. While the use of the cre-loxP system is becoming increasingly widespread, in particular because of growing availability of conditional mouse mutants, many considerations need to be taken into account when utilizing the cre-loxP system. This review provides an overview of the cre-loxP system and its various permutations. It addresses the limitations of cre-loxP technology and related considerations for experimental design, and it discusses alternative strategies for site-specific genetic recombination and integration. © 2017 by John Wiley & Sons, Inc.

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来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.