Ligand-activated site-specific recombination in mice
Ligand-activated site-specific recombination in mice
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DOI：
10.1073/pnas.93.20.10887
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发表时间：
1996-10-01
影响因子：
11.1
通讯作者：
Chambon, P
中科院分区：
文献类型：
--
作者：
Feil, R;Brocard, J;Chambon, P
目前的小鼠基因打靶技术不能在选择的时间和/或在给定的组织中引入体细胞突变，我们在这里报道了使用新版本的Cre/lox系统在小鼠中实现有条件的位点特异性重组，Cre重组酶已与突变的人雌激素受体(ER)的配体结合区域融合，产生了依赖于他莫昔芬的Cre重组酶Cre-ER(T)，该重组酶可被他莫昔芬激活，但不被雌二醇激活。在巨细胞病毒启动子的控制下获得了表达Cre-ER(T)的转基因小鼠。我们的研究表明，在这些转基因小鼠中使用他莫昔芬可以诱导切除位于loxP位点两侧的染色体整合基因，而在未经处理的动物中不能检测到该基因的切除。这种有条件的位点特异性重组系统应该可以分析传统的基因打靶无法解决的敲除表型。
Current mouse gene targeting technology is unable to introduce somatic mutations at a chosen time and/or in a given tissue, We report here that conditional site-specific recombination can be achieved in mice using a new version of the Cre/lox system, The Cre recombinase has been fused to a mutated ligand-binding domain of the human estrogen receptor (ER) resulting in a tamoxifen-dependent Cre recombinase, Cre-ER(T), which is activated by tamoxifen, but not by estradiol. Transgenic mice were generated expressing Cre-ER(T) under the control of a cytomegalovirus promoter. We show that excision of a chromosomally integrated gene flanked by loxP sites can be induced by administration of tamoxifen to these transgenic mice, whereas no excision could be detected in untreated animals. This conditional site-specific recombination system should allow the analysis of knockout phenotypes that cannot be addressed by conventional gene targeting.