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

Nat Biotechnol. 2019 Dec;37(12):1493-1501. doi: 10.1038/s41587-019-0235-7. Epub 2019 Sep 23.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

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

Substances

  • RNA, Guide, CRISPR-Cas Systems