Design of polydactyl zinc-finger proteins for unique addressing within complex genomes
Design of polydactyl zinc-finger proteins for unique addressing within complex genomes
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DOI：
10.1073/pnas.94.11.5525
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发表时间：
1997-05-27
影响因子：
11.1
通讯作者：
Barbas, CF
中科院分区：
文献类型：
--
作者：
Liu, Q;Segal, DJ;Barbas, CF
Cys(2)-His(2)型锌指蛋白是一类具有可塑性的DNA结合蛋白，可被选择结合多种序列。通常，含有三个锌指结构域的锌指蛋白，如鼠转录因子Zif268和人转录因子Sp1，可结合九个连续的碱基对。为了创建一类普遍适用于靶向复杂基因组内独特位点的蛋白质，我们利用基于结构的建模设计了一种连接两个三指蛋白的多肽接头。创建了两种六指蛋白，并证明它们以序列特异性的方式结合18个连续的DNA碱基对。将这些蛋白表达为与激活或抑制结构域的融合体，可使人细胞内的转录特异性上调或下调。多指锌指蛋白应可广泛应用于基因治疗策略以及新型转基因植物和动物的开发中，作为基因组特异性的转录开关。
Zinc-finger proteins of the Cys(2)-His(2) type represent a class of malleable DNA-binding proteins that may be selected to bind diverse sequences, Typically, zinc-finger proteins containing three zinc-finger domains, like the murine transcription factor Zif268 and the human transcription factor Sp1, bind nine contiguous base pairs, To create a class of proteins that would be generally applicable to target unique sites within complex genomes, we have utilized structure-based modeling to design a polypeptide linker that fuses two three-finger proteins, Two six-fingered proteins were created and demonstrated to bind 18 contiguous bp of DNA in a sequence-specific fashion, Expression of these proteins as fusions to activation or repression domains allows transcription to be specifically up- or down-modulated within human cells, Polydactyl zinc-finger proteins should be broadly applicable as genome-specific transcriptional switches in gene therapy strategies and the development of novel transgenic plants and animals.