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

Open access
Luo, Michelle L., Mullis, Adam S., Leenay, Ryan T., Beisel, Chase L.·2015·Nucleic Acids Research

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.

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Metadata

Authors
Luo, M. L., Mullis, A. S., Leenay, R. T., & Beisel, C. L.
Venue
Nucleic Acids Research
Published
2014-10-17
Volume
43
Issue
1
Pages
674-681
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
OpenAlex
W2117444879
Cited by
245
References
14
FWCI
7.30
Type
journal article
Language
en
Open access
gold

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CoChat. (2026). Citation check: Repurposing endogenous type I CRISPR-Cas systems for programmable gene repression. https://app.cochat.ai/papers/10.1093/nar/gku971
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Luo, M. L., Mullis, A. S., Leenay, R. T., & Beisel, C. L. (2015). Repurposing endogenous type I CRISPR-Cas systems for programmable gene repression. Nucleic Acids Research. https://doi.org/10.1093/nar/gku971

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