Comprehensive interrogation of natural TALE DNA-binding modules and transcriptional repressor domains.

Comprehensive interrogation of natural TALE DNA-binding modules and transcriptional repressor domains.
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DOI:
10.1038/ncomms1962
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发表时间:
2012-07-24
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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转录激活因子样效应器(TALE)是序列特异性的DNA结合蛋白,具有模块化、重复的DNA结合域。TALEs已经能够为基因组工程创建可定制的设计转录因子和序列特异性核酸酶。在这里,我们报告了两个改进的TALE工具箱实现有效的激活和抑制内源性基因表达在哺乳动物细胞。我们发现自然存在的重复可变双残基(RVD) Asn-His (NH)对鸟嘌呤具有高的生物活性和特异性,鸟嘌呤是哺乳动物基因组中高度普遍的碱基。我们还报道了一种有效的TALE转录抑制物结构,用于靶向抑制哺乳动物细胞的转录。这些发现将提高使用TALEs可以实现的基因组工程的精确性和有效性。
Transcription activator-like effectors (TALE) are sequence-specific DNA binding proteins that harbor modular, repetitive DNA binding domains. TALEs have enabled the creation of customizable designer transcriptional factors and sequence-specific nucleases for genome engineering. Here we report two improvements of the TALE toolbox for achieving efficient activation and repression of endogenous gene expression in mammalian cells. We show that the naturally occurring repeat variable diresidue (RVD) Asn-His (NH) has high biological activity and specificity for guanine, a highly prevalent base in mammalian genomes. We also report an effective TALE transcriptional repressor architecture for targeted inhibition of transcription in mammalian cells. These findings will improve the precision and effectiveness of genome engineering that can be achieved using TALEs.
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