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

Abstract

Class 2 CRISPR-Cas systems, such as Cas9 and Cas12, have been widely applied for targeting DNA sequences in eukaryotic genomes. However, class I 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 E. coli and L. 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 I CRISPR-Cas systems for further exploration.