Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria

• Published in: Nature communications Vol. 9; no. 1; pp. 2489 - 11
• Main Authors: Dong, Chen, Fontana, Jason, Patel, Anika, Carothers, James M., Zalatan, Jesse G.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 27.06.2018; Nature Publishing Group; Nature Portfolio
• Subjects: 13/31; 42/41; 631/1647/338/552; 631/337/4041; 631/61/318; Bacteria; Biosynthesis; Cellular apoptosis susceptibility protein; Chromatin; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; DNA; E coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Editing; Gene expression; Gene Knockout Techniques; Genes, Synthetic - genetics; Genome, Bacterial - genetics; Humanities and Social Sciences; Kinases; Metabolic Engineering - methods; Metabolic Networks and Pathways - genetics; multidisciplinary; Science; Science (multidisciplinary); Transcription activation; Transcription factors; Transcription Initiation Site; Transcriptional Activation - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-018-04901-6

Methods to regulate gene expression programs in bacterial cells are limited by the absence of effective gene activators. To address this challenge, we have developed synthetic bacterial transcriptional activators in E. coli by linking activation domains to programmable CRISPR-Cas DNA binding domains. Effective gene activation requires target sites situated in a narrow region just upstream of the transcription start site, in sharp contrast to the relatively flexible target site requirements for gene activation in eukaryotic cells. Together with existing tools for CRISPRi gene repression, these bacterial activators enable programmable control over multiple genes with simultaneous activation and repression. Further, the entire gene expression program can be switched on by inducing expression of the CRISPR-Cas system. This work will provide a foundation for engineering synthetic bacterial cellular devices with applications including diagnostics, therapeutics, and industrial biosynthesis. The absence of effective gene activators in bacteria limits regulated expression programs. Here the authors design synthetic bacterial CRISPR-Cas transcriptional activators that can be used to construct multi-gene programs of activation and repression.