Document Type

Article

Publication Date

9-20-2019

Keywords

JGM

JAX Source

Nat Commun 2019 Sep 20; 10(1):4296

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.

Comments

This open access article is licensed under a Creative Commons Attribution 4.0 International License.

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