Document Type
Article
Publication Date
9-20-2019
Keywords
JGM
JAX Source
Nat Commun 2019 Sep 20; 10(1):4296
Volume
10
Issue
1
First Page
4296
Last Page
4296
ISSN
2041-1723
PMID
31541098
DOI
https://doi.org/10.1038/s41467-019-12339-7
Grant
HG009900,CA034196,Leukemia Research Foundation New Investigator Grant,Jackson Laboratory Director's Innovation Fund
Abstract
Here we develop a methylation editing toolbox, Casilio-ME, that enables not only RNA-guided methylcytosine editing by targeting TET1 to genomic sites, but also by co-delivering TET1 and protein factors that couple methylcytosine oxidation to DNA repair activities, and/or promote TET1 to achieve enhanced activation of methylation-silenced genes. Delivery of TET1 activity by Casilio-ME1 robustly alters the CpG methylation landscape of promoter regions and activates methylation-silenced genes. We augment Casilio-ME1 to simultaneously deliver the TET1-catalytic domain and GADD45A (Casilio-ME2) or NEIL2 (Casilio-ME3) to streamline removal of oxidized cytosine intermediates to enhance activation of targeted genes. Using two-in-one effectors or modular effectors, Casilio-ME2 and Casilio-ME3 remarkably boost gene activation and methylcytosine demethylation of targeted loci. We expand the toolbox to enable a stable and expression-inducible system for broader application of the Casilio-ME platforms. This work establishes a platform for editing DNA methylation to enable research investigations interrogating DNA methylomes.
Recommended Citation
Taghbalout A,
Du M,
Jillette NL,
Rosikiewicz W,
Rath A,
Heinen C,
Li S,
Cheng A.
Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nat Commun 2019 Sep 20; 10(1):4296
Comments
This open access article is licensed under a Creative Commons Attribution 4.0 International License.