A Cre recombinase transgene with mosaic, widespread tamoxifen-inducible action
A Cre recombinase transgene with mosaic, widespread tamoxifen-inducible action
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DOI：
10.1002/gene.10021
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发表时间：
2002-01-01
期刊：
影响因子：
1.5
通讯作者：
Lobe, CG
中科院分区：
文献类型：
--
作者：
Guo, CY;Yang, WY;Lobe, CG
Cre介导的位点特异性重组允许在小鼠中进行条件性转基因表达或基因敲除。诱导型Cre重组系统已被开发出来，以绕过最初的胚胎致死表型，并能够研究后期胚胎或成年表型。我们已经培育出Cre转基因小鼠，其切除是他莫昔芬诱导的，并以广泛的嵌合模式发生。我们利用我们的Cre切除报告系统结合胚胎干细胞（ES）细胞筛选，来鉴定在没有他莫昔芬时Cre背景活性不可检测，但添加他莫昔芬后能有效切除的ES细胞克隆。从这些ES细胞衍生的CreER(TM)转基因小鼠品系使用Z/AP和Z/EG Cre报告品系进行了测试。报告基因表达表明，在给予他莫昔芬2天后，Cre切除在妊娠中期胚胎中达到最大值，Cre切除细胞的总体效率为5 - 10%。在他莫昔芬处理3天后，大多数报告基因表达标记细胞群，表明Cre切除细胞的扩增，并且Cre切除细胞的比例得以维持。在成年小鼠中，在他莫昔芬处理后，所有组织中也观察到不同效率的Cre切除。（C）2002威利 - 利斯公司
Cre-mediated site-specific recombination allows conditional transgene expression or gene knockouts in mice. Inducible Cre recombination systems have been developed to bypass initial embryonic lethal phenotypes and provide access to later embryonic or adult phenotypes. We have produced Cre transgenic mice in which excision is tamoxifen inducible and occurs in a widespread mosaic pattern. We utilized our Cre excision reporter system combined with an embryonic stem (ES) cell screen to identify ES cell clones with undetectable background Cre activity in the absence of tamoxifen but efficient excision upon addition of tamoxifen. The CreER(TM) transgenic mouse lines derived from the ES cells were tested using the Z/AP and Z/EG Cre reporter lines. Reporter gene expression indicated Cre excision was maximal in midgestation embryos by 2 days after tamoxifen administration, with an overall efficiency of 5-10% of cells with Cre excision. At 3 days after tamoxifen treatment most reporter gene expression marked groups of cells, suggesting an expansion of cells with Cre excision, and the proportion of cells with Cre excision was maintained. In adults, Cre excision was also observed with varying efficiencies in all tissues after tamoxifen treatment. (C) 2002 Wiley-Liss, Inc.