Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells
Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells
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DOI：
10.1073/pnas.94.26.14602
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发表时间：
1997-12-23
影响因子：
11.1
通讯作者：
Marchuk, Y
中科院分区：
文献类型：
--
作者：
O'Gorman, S;Dagenais, NA;Marchuk, Y
通过结合使用位点特异性和同源重组在小鼠中产生微小或条件突变已经成为哺乳动物遗传学中日益广泛的实验范例。含有重组酶转基因的胚胎干细胞在雄性生殖系中表达，但不在其他组织或胚胎干细胞本身中表达，这将大大简化此类等位基因的产生，在这里，我们显示了由小鼠鱼精蛋白1启动子和Cre重组酶编码序列组成的转基因介导Cre靶转基因在雄性生殖系中的有效重组，但在其他组织中不介导Cre靶转基因的有效重组。用包括loxP侧翼的选择性标记的靶向载体转染从这些转基因株之一产生的胚胎干细胞系，这些结果确立了该系统在小鼠中产生同源重组等位基因所需的时间、努力和资源的潜力，所述同源重组等位基因已被位点特异性重组酶二次重排。
The production of subtle or conditional mutations in mice through the combined use of site-specific and homologous recombination has become an increasingly widespread experimental paradigm in mammalian genetics, Embryonic stem cells containing recombinase transgenes that were expressed in the male germ line, but not in other tissues or in the embryonic stem cells themselves, would substantially simplify the production of such alleles, Here we show that transgenes comprised of the mouse protamine 1 promoter and the Cre recombinase coding sequence mediate the efficient recombination of a Cre target transgene in the male germ line, but not in other tissues, Embryonic stem cell lines generated from one of these transgenic strains were transfected with targeting vectors that included loxP-flanked selectable markers, and homologously recombined alleles containing the marker and functional loxP sites were isolated, These results establish the potential of the system for substantially reducing the time, effort, and resources required to produce homologously recombined alleles in mice that have been secondarily rearranged by a site-specific recombinase.