Jullien Nicolas, Goddard Isabelle, Selmi-Ruby Samia, Fina Jean-Luc, Cremer Harold, Herman Jean-Paul
Genesis. 2008 Apr;46(4):193-9. doi: 10.1002/dvg.20383.
We examined the use of ERT2-iCre-ERT2 (Cre2ERT2), a tamoxifen-regulated form of Cre that has been described to have a background activity lower than that of other tamoxifen-regulated Cre constructs, for establishing performant conditional deleter mouse lines. Cre2ERT2 was inserted by homologous recombination into the Rosa26 locus. These mice were mated with R26R Cre-reporter mice. No recombination could be observed in the progenies in the absence of tamoxifen treatment. Tamoxifen treatment at E13-14 led to a high level, albeit variable, recombination in most of the tissues examined: liver, heart, kidney, brain, lung etc. Treatment of adult animals also induced recombination in these tissues, although at a lower level. Northern blot and qPCR studies suggested that these differences are not linked to significant variations of the level of expression of Cre2ERT2. Thus, Cre2ERT2 appears to be a good alternative to existing modulatable Cre systems, displaying a lack of background activity and a high-level inducibility in vivo.
我们研究了ERT2-iCre-ERT2(Cre2ERT2)的应用,它是一种他莫昔芬调节型Cre,据报道其背景活性低于其他他莫昔芬调节型Cre构建体,用于建立高效的条件性敲除小鼠品系。通过同源重组将Cre2ERT2插入Rosa26基因座。将这些小鼠与R26R Cre报告基因小鼠交配。在未进行他莫昔芬处理的后代中未观察到重组。在E13 - 14期进行他莫昔芬处理后,在大多数检测的组织中,如肝脏、心脏、肾脏、大脑、肺等,均导致了高水平(尽管存在差异)的重组。对成年动物进行处理也能在这些组织中诱导重组,不过水平较低。Northern印迹和qPCR研究表明,这些差异与Cre2ERT2表达水平的显著变化无关。因此,Cre2ERT2似乎是现有可调节Cre系统的一个良好替代方案,在体内表现出缺乏背景活性和高水平诱导性的特点。