Tetracycline repressor, tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible gene expression in mammalian cells
Tetracycline repressor, tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible gene expression in mammalian cells
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DOI：
10.1089/hum.1998.9.13-1939
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发表时间：
1998-09-01
影响因子：
4.2
通讯作者：
Eriksson, E
中科院分区：
文献类型：
--
作者：
Yao, F;Svensjö, T;Eriksson, E
这篇文章描述了第一个(据我们所知)四环素诱导的调控系统，它证明了四环素抑制因子(TetR)本身，而不是TetR-哺乳动物细胞转录因子融合衍生物，可以作为一个强大的反式调节剂来调节哺乳动物细胞中的基因表达。通过正确定位TATA元件下游的四环素操纵子和人表皮生长因子(HEGF)作为报告基因，我们发现四环素操纵子携带HCMV主要立即早期增强子-启动子(PcmvetO)的基因表达可以受到TetR的三个数量级以上的调控，当(1)报告单元在瞬时表达实验中与TetR表达载体共转染时，(2)报告单元稳定整合到TetR表达细胞系的染色体中，这种TetR介导的可诱导基因调控水平显著高于其他基于抑制的哺乳动物细胞转录开关系统。在活体猪创伤模型中，检测到近60倍的TetR介导的调节作用，当给予四环素时，这种调节作用被逆转。总而言之，这项研究提供了一种直接实现这种四环素诱导的调控开关来控制基因在体外、体内和基因治疗中的表达。
This article describes the first (to our knowledge) tetracycline-inducible regulatory system that demonstrates that the tetracycline repressor (tetR) alone, rather than tetR-mammalian cell transcription factor fusion derivatives, can function as a potent trans-modulator to regulate gene expression in mammalian cells. With proper positioning of tetracycline operators downstream of the TATA element and of human epidermal growth factor (hEGF) as a reporter, we show that gene expression from the tetracycline operator-bearing hCMV major immediate-early enhancer-promoter (pcmvtetO) can be regulated by tetR over three orders of magnitude in response to tetracycline when (1) the reporter was cotransfected with tetR-expressing plasmid in transient expression assays, and (2) the reporter unit was stably integrated into the chromosome of a tetR-expressing cell line, This level of tetR-mediated inducible gene regulation is significantly higher than that of other repression-based mammalian cell transcription switch systems. In an in vivo porcine wound model, close to 60-fold tetR-mediated regulatory effects were detected and it was reversed when tetracycline was administered. Collectively, this study provides a direct implementation of this tetracycline-inducible regulatory switch for controlling gene expression in vitro, in vivo, and in gene therapy.