Stec D E, Sigmund C D
Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa, USA.
Exp Nephrol. 1998 Nov-Dec;6(6):568-75. doi: 10.1159/000020573.
With the advent of gene-targeting in mouse embryonic stem (ES) cells, the use of knockout mice to study the physiological effects of loss of gene function has become increasingly prevalent. However, there are several drawbacks with conventional gene-targeting approaches which may make phenotyping of the resultant mice difficult, if not, impossible. Conventional gene-targeting results in the loss of function of the targeted gene in all cells and tissues, which can be problematic for genes which are required developmentally, which exhibit a wide tissue-specific expression pattern, or are part of complex paracrine systems. As with mice that lack the angiotensinogen or endothelin-1 gene, loss of gene function may lead to a lethal phenotype which can be manifested during embryonic development, at birth or postnatally. These limitations could potentially be circumvented by using a system in which the loss of gene function is placed under spatial and/or temporal control. We will discuss how the cre-loxP recombinase system can be applied to delete a gene in a tissue- and developmentally regulated fashion.
随着小鼠胚胎干细胞基因靶向技术的出现，利用基因敲除小鼠来研究基因功能缺失的生理效应变得越来越普遍。然而，传统的基因靶向方法存在一些缺点，这些缺点可能会使对所得小鼠进行表型分析变得困难，甚至无法进行。传统的基因靶向会导致所有细胞和组织中靶向基因功能的丧失，对于那些在发育过程中必需、表现出广泛的组织特异性表达模式或属于复杂旁分泌系统一部分的基因来说，这可能会产生问题。就像缺乏血管紧张素原或内皮素 -1 基因的小鼠一样，基因功能的丧失可能会导致致死表型，这种表型可能在胚胎发育期间、出生时或出生后表现出来。通过使用一种能将基因功能丧失置于空间和 / 或时间控制之下的系统，这些限制有可能被规避。我们将讨论如何应用 cre-loxP 重组酶系统以组织和发育调控的方式删除基因。
