A strategy for enhancing the transcriptional activity of weak cell type-specific promoters
A strategy for enhancing the transcriptional activity of weak cell type-specific promoters
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DOI：
10.1038/sj.gt.3300778
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发表时间：
1998-12-01
期刊：
影响因子：
5.1
通讯作者：
Müller, R
中科院分区：
文献类型：
--
作者：
Nettelbeck, DM;Jérôme, V;Müller, R
细胞类型和组织特异性启动子在基因治疗的位点选择性载体的开发中起着重要作用。已经描述了大量的高度特异性启动子，但是它们的适用性经常受到它们低效的转录活性的阻碍。在这项研究中，我们描述了一种新的策略，增强弱启动子的活性，而不丧失特异性。该策略的基本原理是建立一个由细胞类型特异性启动子转录启动的正反馈环。这是通过使用细胞类型特异性启动子来驱动所需效应子/报告基因产物的同时表达和强人工转录激活因子来实现的，所述强人工转录激活因子通过启动子中的适当结合位点刺激转录。使用VP 16-莱克萨嵌合转录因子，我们表明，这种方法导致14-> 100倍的增强和胃肠道特异性蔗糖酶异麦芽糖酶启动子，同时保持约30-> 100倍的细胞类型特异性。
Cell type- and tissue-specific promoters play an important role in the development of site-selective vectors for gene therapy. A large number of highly specific promoters has been described, but their applicability is often hampered by their inefficient transcriptional activity. In this study, we describe a new strategy for enhancing the activity of weak promoters without loss of specificity. The basic principle of this strategy is to establish a positive feedback loop which is initiated by transcription from a cell type-specific promoter. This was achieved by using a cell type-specific promoter to drive the simultaneous expression of the desired effector/reporter gene product and a strong artificial transcriptional activator which stimulates transcription through appropriate binding sites in the promoter. Using a VP16-LexA chimeric transcription factor, we show that this approach leads to a 14- to > 100-fold enhancement of both and the gastrointestinal-specific sucrase-isomaltase promoter while maintaining approximately 30- to > 100-fold cell type specificity.