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Cre驱动小鼠品系的表征与验证

Characterization and Validation of Cre-Driver Mouse Lines.

作者信息

Gofflot Françoise, Wendling Olivia, Chartoire Nathalie, Birling Marie-Christine, Warot Xavier, Auwerx Johan

机构信息

Institut Clinique de la Souris (ICS), Illkirch, France.

Université Catholique de Louvain, Life Science Institute, Louvain-la-Neuve, Belgium.

出版信息

Curr Protoc Mouse Biol. 2011 Mar 1;1(1):1-15. doi: 10.1002/9780470942390.mo100103.

Abstract

Conditional gene manipulations in mice are increasingly popular strategies in biomedical research. These approaches rely on the production of conditional genetically engineered mutant mouse (GEMM) lines with mutations in protein-encoding genes. These conditional GEMMs are then bred with one or several transgenic mouse lines expressing a site-specific recombinase, most often the Cre recombinase, in a tissue-specific manner. Conditional GEMMs can only be exploited if Cre transgenic mouse lines are available to generate somatic mutations, and thus the number of Cre transgenic lines has significantly increased over the last 15 years. Once produced, these transgenic lines must be validated for reliable, efficient, and specific Cre expression and Cre-mediated recombination. In this overview, the minimum level of information that is ideally required to validate a Cre-driver transgenic line is first discussed. The vagaries associated with validation procedures are considered next, and some solutions are proposed to assess the expression and activity of constitutive or inducible Cre recombinase before undertaking extensive breeding experiments and exhaustive phenotyping. Curr. Protoc. Mouse Biol. 1:1-15. © 2011 by John Wiley & Sons, Inc.

摘要

在生物医学研究中,小鼠条件性基因操作是越来越流行的策略。这些方法依赖于构建条件性基因工程突变小鼠(GEMM)品系,其蛋白质编码基因存在突变。然后将这些条件性GEMM与一个或几个以组织特异性方式表达位点特异性重组酶(最常见的是Cre重组酶)的转基因小鼠品系进行杂交。只有当Cre转基因小鼠品系可用于产生体细胞突变时,条件性GEMM才能发挥作用,因此在过去15年中,Cre转基因品系的数量显著增加。一旦产生,这些转基因品系必须经过验证,以确保Cre表达可靠、高效且具有特异性,以及Cre介导的重组有效。在本综述中,首先讨论了验证Cre驱动转基因品系理想情况下所需的最低信息量。接下来考虑与验证程序相关的变幻莫测的因素,并提出一些解决方案,以便在进行广泛的杂交实验和详尽的表型分析之前评估组成型或诱导型Cre重组酶的表达和活性。《小鼠生物学实验指南》第1卷:1 - 15页。© 2011年约翰·威利父子公司版权所有。

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