A highly efficient ligand-regulated Cre recombinase mouse line shows that LoxP recombination is position dependent
A highly efficient ligand-regulated Cre recombinase mouse line shows that LoxP recombination is position dependent
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DOI：
10.1093/embo-reports/kve064
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发表时间：
2001-04-01
期刊：
影响因子：
7.7
通讯作者：
Berns, A
中科院分区：
文献类型：
--
作者：
Vooijs, M;Jonkers, J;Berns, A
利用Cre/loxP系统进行的条件性基因失活被广泛应用，但对Cre表达进行恰当调控的困难仍然是瓶颈之一。调控Cre活性的一种方法是利用一种突变的雌激素激素结合结构域（ERT）使Cre保持无活性，除非存在非甾体类雌激素类似物4 - 羟基他莫昔芬（OHT）。在此我们描述了一种从广泛表达的ROSA26（R26）基因座表达Cre - ERT的小鼠品系。我们证明了在体内以及源自这些小鼠的原代细胞中Cre重组的高效时空调控。我们表明在同一细胞内不同底物之间重组频率存在显著差异。当需要在同一细胞内同时转换多个等位基因时，这会产生重要影响，并凸显了使用报告基因小鼠作为指示品系时可能遇到的困难之一。
Conditional gene inactivation using the Cre/loxP system is widely used, but the difficulty in properly regulating Cre expression remains one of the bottlenecks. One approach to regulate Cre activity utilizes a mutant estrogen hormone-binding domain (ERT) to keep Cre inactive unless the nonsteroidal estrogen analog 4-hydroxytamoxifen (OHT) is present. Here we describe a mouse strain expressing Cre-ERT from the ubiquitously expressed ROSA26 (R26) locus. We demonstrate efficient temporal and spatial regulation of Cre recombination in vivo and in primary cells derived from these mice. We show the existence of marked differences in recombination frequencies between different substrates within the same cell. This has important consequences when concurrent switching of multiple alleles within the same cell is needed, and highlights one of the difficulties that may be encountered when using reporter mice as indicator strains.