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

• Published in: Nature communications Vol. 3; no. 1; p. 968
• Main Authors: Cong, Le, Zhou, Ruhong, Kuo, Yu-chi, Cunniff, Margaret, Zhang, Feng
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24.07.2012; Nature Publishing Group
• Subjects: 631/208/200; 631/337/572; 631/45/535; 631/45/612/1229; Cell Line; Computational Biology - methods; DNA - chemistry; DNA - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Genetic Engineering - methods; Humanities and Social Sciences; Humans; Models, Molecular; multidisciplinary; Science; Science (multidisciplinary); Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - metabolism; Transcriptional Activation - genetics; Transcriptional Activation - physiology
• ISSN: 2041-1723; 2041-1723
• 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. The peptide sequence of transcription activator-like effectors (TALEs) can be customized to tailor the binding of TALEs to specific DNA sequences. Cong et al . improve TALE specificity for guanine binding and use a genetic construct based on TALEs to efficiently repress expression of a target gene.