Transgenic mice for in vivo epigenome editing with CRISPR-based systems
- PMID: 34341582
- PMCID: PMC8349887
- DOI: 10.1038/s41592-021-01207-2
Transgenic mice for in vivo epigenome editing with CRISPR-based systems
Abstract
CRISPR-Cas9 technologies have dramatically increased the ease of targeting DNA sequences in the genomes of living systems. The fusion of chromatin-modifying domains to nuclease-deactivated Cas9 (dCas9) has enabled targeted epigenome editing in both cultured cells and animal models. However, delivering large dCas9 fusion proteins to target cells and tissues is an obstacle to the widespread adoption of these tools for in vivo studies. Here, we describe the generation and characterization of two conditional transgenic mouse lines for epigenome editing, Rosa26:LSL-dCas9-p300 for gene activation and Rosa26:LSL-dCas9-KRAB for gene repression. By targeting the guide RNAs to transcriptional start sites or distal enhancer elements, we demonstrate regulation of target genes and corresponding changes to epigenetic states and downstream phenotypes in the brain and liver in vivo, and in T cells and fibroblasts ex vivo. These mouse lines are convenient and valuable tools for facile, temporally controlled, and tissue-restricted epigenome editing and manipulation of gene expression in vivo.
© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.
Conflict of interest statement
Conflict of Interest Statement
CAG, IBH, and TER have filed patent applications related to CRISPR technologies for genome engineering. CAG is an advisor to Tune Therapeutics, Sarepta Therapeutics, Levo Therapeutics, and Iveric Bio, and a co-founder of Tune Therapeutics, Element Genomics, and Locus Biosciences. AA is a co-founder and advisor to StrideBio and TorqueBio. TER is a co-founder of Element Genomics. MPG is a co-founder and employee of Tune Therapeutics. All other authors declare no conflicts of interest.
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