Transcriptional and posttranslational regulation of Cre recombinase by RU486 as the basis for an enhanced inducible expression system.
Transcriptional and posttranslational regulation of Cre recombinase by RU486 as the basis for an enhanced inducible expression system.
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RU486 对 Cre 重组酶的转录和翻译后调节作为增强诱导表达系统的基础。
DOI：
10.1016/j.ymthe.2003.07.005
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Federoff,HowardJ
中科院分区：
文献类型：
--
作者：
Kyrkanides,Stephanos;Miller,JennieH;Bowers,WilliamJ;Federoff,HowardJ
遗传学研究通常需要使用诱导型表达系统，由此特定基因的表达可以通过外源施用惰性配体来调节。基于Cre/loxP的系统先前已被描述为通过施加位点特异性DNA重组的诱导型表达系统的基础。在我们的努力，以提高RU 486响应CrePr 1构建的属性，我们已经开发了双GLVP/CrePr系统，其中RU 486赋予活性控制在转录和翻译后水平的CrePr 1。这是通过将CrePr 1转录调控置于RU 486敏感的嵌合调节剂GLVP的控制下来实现的。开发了具有双重GLVP/CrePr以及单一CrePr 1系统的稳定细胞系。我们的研究结果表明，双重调节系统是高度可诱导的RU 486，同时保持最小的基础活动（“泄漏”），特征，可用于转基因小鼠的发展，其中遗传途径可以打开或关闭后，外源性管理RU 486在生理惰性剂量。
Genetic studies often require the employment of an inducible expression system, whereby the expression of a particular gene can be regulated by the exogenous administration of an inert ligand. Cre/loxP-based systems have been previously described as the basis for inducible expression systems by exerting site-specific DNA recombination. In our effort to enhance the properties of the RU486-responsive CrePr1 construct, we have developed the dual GLVP/CrePr system, in which RU486 confers activity control at both the transcriptional and the posttranslational level of CrePr1. This was achieved by placing CrePr1 transcriptional regulation under the control of the RU486-sensitive chimeric regulator GLVP. Stable cell lines harboring the dual GLVP/CrePr as well as the single CrePr1 system were developed. Our results indicate that the dually regulated system is highly inducible by RU486 while maintaining minimal basal activity ("leakage"), characteristics that can be employed in the development of transgenic mice in which genetic pathways can be turned on or turned off after exogenous administration of RU486 at physiologically inert doses.