Format

Send to

Choose Destination
Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.

Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria.

Author information

1
Department of Chemistry, University of Washington, Seattle, WA, 98195, USA.
2
Molecular Engineering & Sciences Institute, University of Washington, Seattle, WA, 98195, USA.
3
Molecular Engineering & Sciences Institute, University of Washington, Seattle, WA, 98195, USA. jcaroth@uw.edu.
4
Department of Chemical Engineering, University of Washington, Seattle, WA, 98195, USA. jcaroth@uw.edu.
5
Center for Synthetic Biology, University of Washington, Seattle, WA, 98195, USA. jcaroth@uw.edu.
6
Department of Chemistry, University of Washington, Seattle, WA, 98195, USA. zalatan@uw.edu.
7
Molecular Engineering & Sciences Institute, University of Washington, Seattle, WA, 98195, USA. zalatan@uw.edu.
8
Center for Synthetic Biology, University of Washington, Seattle, WA, 98195, USA. zalatan@uw.edu.

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.

PMID:
29950558
PMCID:
PMC6021436
DOI:
10.1038/s41467-018-04901-6
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center