A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities
A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities
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DOI：
10.1038/sj.gt.3301443
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发表时间：
2001-05-01
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：
Jérôme, V;Müller, R
基因治疗面临的最大挑战之一是开发将治疗基因的活性特异性地导向疾病部位的载体。为了实现这一目标，通过具有多种特异性的合成启动子限制转基因转录代表了特别有前途的策略。为此，我们开发了一种普遍适用的策略（DCTF系统），其中合成启动子由人工异二聚体转录因子驱动，该转录因子的DNA结合和反式激活亚基从具有不同选择性的两个启动子表达。DCTF系统的一个关键决定因素是异源二聚化界面，其应提供高亲和力相互作用而不受内源性蛋白质的干扰。在这里，我们描述了这样一个二聚化系统的基础上工程Fos和Jun亮氨酸拉链。我们展示了该系统的有用性，细胞类型特异性和细胞周期调节转录的组合，并证明了其在体内环境中的功能。
One of the biggest challenges facing gene therapy is the development of Vectors that direct the activity of therapeutic genes specifically to the sites of disease. To achieve this goal, the restriction of transgene transcription via synthetic promoters that are endowed with multiple specificities represents a particularly promising strategy. Towards this end, we have developed a generally applicable strategy (DCTF system) where a synthetic promoter is driven by an artificial heterodimeric transcription factor whose DNA-binding and transactivating subunits are expressed from two promoters with different selectivity. A crucial determinant of the DCTF system is the heterodimerization interface that should provide for a high affinity interaction without interference by endogenous proteins. Here, we describe such a dimerization system based on engineered Fos and Jun leucine zippers. We show the usefulness of this system for the combination of cell type-specific and cell cycle-regulated transcription and demonstrate its functionality in an in vivo setting.