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Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells.

Publication ,  Conference
Pickar-Oliver, A; Black, JB; Lewis, MM; Mutchnick, KJ; Klann, TS; Gilcrest, KA; Sitton, MJ; Nelson, CE; Barrera, A; Bartelt, LC; Reddy, TE ...
Published in: Nat Biotechnol
December 2019

Class 2 CRISPR-Cas systems, such as Cas9 and Cas12, have been widely used to target DNA sequences in eukaryotic genomes. However, class 1 CRISPR-Cas systems, which represent about 90% of all CRISPR systems in nature, remain largely unexplored for genome engineering applications. Here, we show that class 1 CRISPR-Cas systems can be expressed in mammalian cells and used for DNA targeting and transcriptional control. We repurpose type I variants of class 1 CRISPR-Cas systems from Escherichia coli and Listeria monocytogenes, which target DNA via a multi-component RNA-guided complex termed Cascade. We validate Cascade expression, complex formation and nuclear localization in human cells, and demonstrate programmable CRISPR RNA (crRNA)-mediated targeting of specific loci in the human genome. By tethering activation and repression domains to Cascade, we modulate the expression of targeted endogenous genes in human cells. This study demonstrates the use of Cascade as a CRISPR-based technology for targeted eukaryotic gene regulation, highlighting class 1 CRISPR-Cas systems for further exploration.

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Published In

Nat Biotechnol

DOI

EISSN

1546-1696

Publication Date

December 2019

Volume

37

Issue

12

Start / End Page

1493 / 1501

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • RNA, Guide, CRISPR-Cas Systems
  • Listeria monocytogenes
  • Humans
  • HEK293 Cells
  • Genetic Engineering
  • Escherichia coli
  • CRISPR-Cas Systems
 

Citation

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Chicago
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MLA
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Pickar-Oliver, A., Black, J. B., Lewis, M. M., Mutchnick, K. J., Klann, T. S., Gilcrest, K. A., … Gersbach, C. A. (2019). Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. In Nat Biotechnol (Vol. 37, pp. 1493–1501). United States. https://doi.org/10.1038/s41587-019-0235-7
Pickar-Oliver, Adrian, Joshua B. Black, Mae M. Lewis, Kevin J. Mutchnick, Tyler S. Klann, Kylie A. Gilcrest, Madeleine J. Sitton, et al. “Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells.” In Nat Biotechnol, 37:1493–1501, 2019. https://doi.org/10.1038/s41587-019-0235-7.
Pickar-Oliver A, Black JB, Lewis MM, Mutchnick KJ, Klann TS, Gilcrest KA, et al. Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. In: Nat Biotechnol. 2019. p. 1493–501.
Pickar-Oliver, Adrian, et al. “Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells.Nat Biotechnol, vol. 37, no. 12, 2019, pp. 1493–501. Pubmed, doi:10.1038/s41587-019-0235-7.
Pickar-Oliver A, Black JB, Lewis MM, Mutchnick KJ, Klann TS, Gilcrest KA, Sitton MJ, Nelson CE, Barrera A, Bartelt LC, Reddy TE, Beisel CL, Barrangou R, Gersbach CA. Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. Nat Biotechnol. 2019. p. 1493–1501.

Published In

Nat Biotechnol

DOI

EISSN

1546-1696

Publication Date

December 2019

Volume

37

Issue

12

Start / End Page

1493 / 1501

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • RNA, Guide, CRISPR-Cas Systems
  • Listeria monocytogenes
  • Humans
  • HEK293 Cells
  • Genetic Engineering
  • Escherichia coli
  • CRISPR-Cas Systems