Codon-improved Cre recombinase (iCre) expression in the mouse
Codon-improved Cre recombinase (iCre) expression in the mouse
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DOI：
10.1002/gene.10023
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发表时间：
2002-01-01
期刊：
影响因子：
1.5
通讯作者：
Sprengel, R
中科院分区：
文献类型：
--
作者：
Shimshek, DR;Kim, J;Sprengel, R
通过将哺乳动物密码子的使用应用于Cre重组酶，我们通过免疫印迹和功能分析确定了Cre在三个不同的哺乳动物细胞系中的表达。改进的Cre(ICre)基因还旨在降低原核编码序列中的高CpG含量，从而减少哺乳动物表观遗传沉默的机会。获得了频率较高的iCre转基因小鼠，并在所有表达iCre的细胞中观察到loxP介导的DNA重组。此外，iCre融合到两个雌激素受体激素结合域以时间控制Cre，活性也可以在转基因小鼠中表达。然而，他莫昔芬诱导的Cre仅产生较低的Cre活性。因此，尽管Cre融合蛋白在大脑中的有效激活需要进一步改进，但我们的研究表明，ICre应该有助于在小鼠中进行遗传实验。(C)2002年Wiley-Liss，Inc.
By applying the mammalian codon usage to Cre recombinase, we improved Cre expression, as determined by immunoblot and functional analysis, in three different mammalian cell lines. The improved Cre (iCre) gene was also designed to reduce the high CpG content of the prokaryotic coding sequence, thereby reducing the chances of epigenetic silencing in mammals. Transgenic iCre expressing mice were obtained with good frequency, and in these mice loxP-mediated DNA recombination was observed in all cells expressing iCre. Moreover, iCre fused to two estrogen receptor hormone binding domains for temporal control of Cre, activity could also be expressed in transgenic mice. However, Cre induction after administration of tamoxifen yielded only low Cre activity. Thus, whereas efficient activation of Cre fusion proteins in the brain needs further improvements, our studies indicate that iCre should facilitate genetic experiments in the mouse. (C) 2002 Wiley-Liss, Inc.