Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA.
Nat Commun. 2012 Jul 24;3:968. doi: 10.1038/ncomms1962.
Transcription activator-like effectors are sequence-specific DNA-binding proteins that harbour modular, repetitive DNA-binding domains. Transcription activator-like effectors have enabled the creation of customizable designer transcriptional factors and sequence-specific nucleases for genome engineering. Here we report two improvements of the transcription activator-like effector toolbox for achieving efficient activation and repression of endogenous gene expression in mammalian cells. We show that the naturally occurring repeat-variable diresidue Asn-His (NH) has high biological activity and specificity for guanine, a highly prevalent base in mammalian genomes. We also report an effective transcription activator-like effector 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 transcription activator-like effectors.
转录激活子样效应物是序列特异性 DNA 结合蛋白，具有模块化、重复的 DNA 结合结构域。转录激活子样效应物使定制设计的转录因子和序列特异性核酸酶的创建成为可能，用于基因组工程。在这里，我们报告了转录激活子样效应物工具箱的两项改进，以实现哺乳动物细胞中内源基因表达的高效激活和抑制。我们表明，天然存在的重复可变二残基天冬酰胺-组氨酸 (NH) 具有针对鸟嘌呤的高生物活性和特异性，鸟嘌呤是哺乳动物基因组中非常常见的碱基。我们还报告了一种有效的转录激活子样效应物转录抑制剂结构，用于靶向抑制哺乳动物细胞中的转录。这些发现将提高使用转录激活子样效应物实现的基因组工程的精度和有效性。
