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

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 c...

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Bibliographic Details
Published in:Nature biotechnology 2019-12, Vol.37 (12), p.1493-1501
Main Authors: Pickar-Oliver, Adrian, Black, Joshua B, Lewis, Mae M, Mutchnick, Kevin J, Klann, Tyler S, Gilcrest, Kylie A, Sitton, Madeleine J, Nelson, Christopher E, Barrera, Alejandro, Bartelt, Luke C, Reddy, Timothy E, Beisel, Chase L, Barrangou, Rodolphe, Gersbach, Charles A
Format: Article
Language:English
Subjects:
Cells
Complex formation
CRISPR
CRISPR-Cas Systems - genetics
Deoxyribonucleic acid
DNA
DNA sequencing
Domains
E coli
Escherichia coli
Escherichia coli - genetics
Gene expression
Gene regulation
Gene sequencing
Genes
Genetic aspects
Genetic Engineering - methods
Genetic transcription
Genome editing
Genomes
Genomics
HEK293 Cells
Humans
International economic relations
Listeria
Listeria monocytogenes
Listeria monocytogenes - genetics
Localization
Mammalian cells
Nucleotide sequence
Nucleotide sequencing
Ribonucleic acid
RNA
RNA, Guide, CRISPR-Cas Systems - genetics
Scientific equipment and supplies industry
Technology application
Tethering
Transcription
Transcription, Genetic - genetics
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Summary: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.
ISSN:1087-0156
1546-1696
DOI:10.1038/s41587-019-0235-7