Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria

Chen Dong, Jason Fontana, Anika Patel, James M. Carothers () and Jesse G. Zalatan ()
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Chen Dong: University of Washington
Jason Fontana: University of Washington
Anika Patel: University of Washington
James M. Carothers: University of Washington
Jesse G. Zalatan: University of Washington

Nature Communications, 2018, vol. 9, issue 1, 1-11

Abstract: Abstract 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.

Date: 2018
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DOI: 10.1038/s41467-018-04901-6

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