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Nucleic Acids Res. 2015 Jan;43(1):674-81. doi: 10.1093/nar/gku971. Epub 2014 Oct 17.

Repurposing endogenous type I CRISPR-Cas systems for programmable gene repression.

Author information

1
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA.
2
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA cbeisel@ncsu.edu.

Abstract

CRISPR-Cas systems have shown tremendous promise as heterologous tools for genome editing and transcriptional regulation. Because these RNA-directed immune systems are found in most prokaryotes, an opportunity exists to harness the endogenous systems as convenient tools in these organisms. Here, we report that the Type I-E CRISPR-Cas system in Escherichia coli can be co-opted for programmable transcriptional repression. We found that deletion of the signature cas3 gene converted this immune system into a programmable gene regulator capable of reversible gene silencing of heterologous and endogenous genes. Targeting promoter regions yielded the strongest repression, whereas targeting coding regions showed consistent strand bias. Furthermore, multi-targeting CRISPR arrays could generate complex phenotypes. This strategy offers a simple approach to convert many endogenous Type I systems into transcriptional regulators, thereby expanding the available toolkit for CRISPR-mediated genetic control while creating new opportunities for genome-wide screens and pathway engineering.

PMID:
25326321
PMCID:
PMC4288209
DOI:
10.1093/nar/gku971
[Indexed for MEDLINE]
Free PMC Article

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