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Cre转基因小鼠品系。

Cre transgenic mouse lines.

作者信息

Wang Xin

机构信息

Faculty of Life Sciences, University of Manchester, Core Technology Facility, Manchester, UK.

出版信息

Methods Mol Biol. 2009;561:265-73. doi: 10.1007/978-1-60327-019-9_17.

Abstract

With the development of the Cre-LoxP system, conditional gene targeting has rapidly become a powerful technology that facilitates the study of gene function. This advanced technique circumvents three major concerns sometimes levelled against conventional transgenic and gene-targeting approaches. First of all, gene ablation may exert its effect in multiple cell and tissue types, creating a complex phenotype in which it is difficult to distinguish direct function in a particular tissue from secondary effects resulting from altered gene function in other tissues. Secondly, a gene deletion expressed in the germ line may cause embryonic lethality, thereby precluding analysis of gene function in the adult tissues. Thirdly, the transgenic approach represents a somewhat surreal over-expression of a given protein often causing spurious phenotypes. The generation of conditional knockout mice is a multiple-step process, which involves mating the flox mutant mouse line (essential exon/s of the gene of interest are flanked by two LoxP sites) and the Cre-expressing mouse line. Over the past few years many inducible and/or tissue-specific Cre mouse lines have been developed. This chapter will give a brief review of the generation of Cre-expressing mouse lines and will discuss the strategy of using these Cre lines. In addition, information regarding established Cre-expressing mouse lines will be provided.

摘要

随着Cre-LoxP系统的发展,条件性基因打靶迅速成为一项强大的技术,有助于基因功能的研究。这项先进技术规避了有时针对传统转基因和基因打靶方法提出的三个主要问题。首先,基因敲除可能在多种细胞和组织类型中发挥作用,产生复杂的表型,在这种表型中,很难将特定组织中的直接功能与其他组织中基因功能改变产生的次要效应区分开来。其次,在生殖系中表达的基因缺失可能导致胚胎致死,从而排除了对成年组织中基因功能的分析。第三,转基因方法代表了给定蛋白质的某种超现实过度表达,常常导致虚假的表型。条件性敲除小鼠的产生是一个多步骤过程,涉及将flox突变小鼠品系(感兴趣基因的必需外显子被两个LoxP位点侧翼包围)与表达Cre的小鼠品系交配。在过去几年中,已经开发出许多可诱导和/或组织特异性的Cre小鼠品系。本章将简要回顾表达Cre的小鼠品系的产生,并讨论使用这些Cre品系的策略。此外,还将提供有关已建立的表达Cre的小鼠品系的信息。

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