Transgenic mice for in vivo epigenome editing with CRISPR-based systems.

TitleTransgenic mice for in vivo epigenome editing with CRISPR-based systems.
Publication TypeJournal Article
Year of Publication2021
AuthorsMP Gemberling, K Siklenka, E Rodriguez, KR Tonn-Eisinger, A Barrera, F Liu, A Kantor, L Li, V Cigliola, MF Hazlett, CA Williams, LC Bartelt, VJ Madigan, JC Bodle, H Daniels, DC Rouse, IB Hilton, A Asokan, M Ciofani, KD Poss, TE Reddy, AE West, and CA Gersbach
JournalNat Methods
Volume18
Start Page965
Issue8
Pagination965 - 974
Date Published08/2021
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.

DOI10.1038/s41592-021-01207-2
Short TitleNat Methods