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

Published

Conference Paper

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.

Full Text

Duke Authors

Cited Authors

  • 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

Published Date

  • December 2019

Published In

Volume / Issue

  • 37 / 12

Start / End Page

  • 1493 - 1501

PubMed ID

  • 31548729

Pubmed Central ID

  • 31548729

Electronic International Standard Serial Number (EISSN)

  • 1546-1696

International Standard Serial Number (ISSN)

  • 1087-0156

Digital Object Identifier (DOI)

  • 10.1038/s41587-019-0235-7