Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription.

Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription.
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DOI:
10.1038/nbt.1775
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发表时间:
2011-02
影响因子:
46.9
通讯作者:
--
中科院分区:
工程技术1区
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将功能结构域定向到特定DNA序列的能力是研究和工程化生物过程的长期追求的目标。来自黄单胞菌属的转录激活因子样效应子(TALE)为设计序列特异性DNA结合蛋白提供了一个有希望的平台。在这里,我们描述了一个强大的和快速的方法,克服了构建TALE重复结构域的困难。我们合成了17个设计师TALE(dTALE),它们被定制为识别特定的DNA结合位点,并证明dTALE可以特异性地调节人类细胞中天然基因组的内源性基因(Sox 2和Klf 4)的转录。dTALE为生物系统的询问和工程化提供了可设计的DNA靶向平台。
The ability to direct functional domains to specific DNA sequences is a long sought-after goal for studying and engineering biological processes. Transcription activator like effectors (TALEs) from Xanthomonas sp. present a promising platform for designing sequence-specific DNA binding proteins. Here we describe a robust and rapid method for overcoming the difficulty of constructing TALE repeat domains. We synthesized 17 designer TALEs (dTALEs) that are customized to recognize specific DNA binding sites, and demonstrate that dTALEs can specifically modulate transcription of endogenous genes (Sox2 and Klf4) from the native genome in human cells. dTALEs provide a designable DNA targeting platform for the interrogation and engineering of biological systems.
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