Mifepristone-inducible LexPR system to drive and control gene expression in transgenic zebrafish
Mifepristone-inducible LexPR system to drive and control gene expression in transgenic zebrafish
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DOI：
10.1016/j.ydbio.2008.04.042
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发表时间：
2008-08-01
影响因子：
2.7
通讯作者：
Parinov, Serguei
中科院分区：
文献类型：
--
作者：
Emelyanov, Alexander;Parinov, Serguei
有效的转基因方法已经成功地应用于包括斑马鱼在内的许多生物体中。然而，转基因表达的精确时空控制仍然难以实现。在这里，我们描述了一个系统的化学诱导基因表达，并证明其可行性，在斑马鱼中产生转基因驱动线。该系统的关键元件是一种杂合转录因子，该因子通过融合细菌莱克萨阻遏物的DNA结合结构域、人黄体酮受体的截短配体结合结构域和人NF-κ B/p65蛋白的激活结构域而工程化。该杂合转录因子（LexPR反式激活因子）与合成类固醇米非司酮（RU-486）结合，并以配体依赖性方式发挥作用，以诱导置于含有莱克萨结合位点的合成操纵子-启动子序列控制下的基因的表达。转基因表达受到严格控制，可以在生命周期的任何阶段通过在水中施用米非司酮来诱导。为了证明该系统的实用性，我们产生了稳定的转基因株系，其允许激活的K-ras（V12）的诱导型组织特异性表达。LexPR表达系统与Ac/Ds介导的转基因技术相结合，在遗传学和生物技术领域具有广泛的应用前景。(C)2008年爱思唯尔公司All rights reserved.
effective transgenesis methods have been successfully employed in Many organisms including zebrafish. However accurate spatiotemporal control of transgene expression is still difficult to achieve. Here we describe a system for chemical-inducible gene expression and demonstrate its feasibility for generating transgenic driver lines in zebrafish. The key element of this system is a hybrid transcription factor engineered by fusion of the DNA-binding domain of the bacterial LexA repressor a truncated ligand-binding domain of the human progesterone receptor and the activation domain of the human NF-kappa B/p65 protein. This hybrid transcription factor (LexPR transactivator) binds to the synthetic steroid mifepristone (RU-486) and functions in a ligind-dependent manner to induce expression of the gene(s) placed under the control of a synthetic operator-promoter sequence that harbors LexA binding sites. Transgene expression is strictly controlled and can be induced at any stage of the life cycle through administration of mifepristone in the water. To demonstrate the utility of this system we generated stable transgenic lines which allow inducible tissue-specific expression of activated K-ras(V12). Combined with the Ac/Ds-mediated transgenesis the LexPR expression system has many potential applications in the fields of genetics and biotechnology. (C) 2008 Elsevier Inc. All rights reserved.