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Sci Rep. 2017 Feb 9;7:42047. doi: 10.1038/srep42047.

Epigenetic Editing of Ascl1 Gene in Neural Stem Cells by Optogenetics.

Author information

  • 1Department of Anatomy &Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA.
  • 2Bindley Bioscience Center, Department of Agricultural &Biological Engineering, Purdue University, West Lafayette, IN, USA.
  • 3Stark Institute of Neuroscience Research, Indiana University School of Medicine, Indianapolis, IN, USA.

Abstract

Enzymes involved in epigenetic processes such as methyltransferases or demethylases are becoming highly utilized for their persistent DNA or histone modifying efficacy. Herein, we have developed an optogenetic toolbox fused to the catalytic domain (CD) of DNA-methyltransferase3A (DNMT3A-CD) or Ten-Eleven Dioxygenase-1 (TET1-CD) for loci-specific alteration of the methylation state at the promoter of Ascl1 (Mash1), a candidate proneuron gene. Optogenetical protein pairs, CRY2 linked to DNMT3A-CD or TET1-CD and CIB1 fused to a Transcription Activator-Like Element (TALE) locating an Ascl1 promoter region, were designed for site specific epigenetic editing. A differentially methylated region at the Ascl1 promoter, isolated from murine dorsal root ganglion (hypermethylated) and striated cells (hypomethylated), was targeted with these optogenetic-epigenetic constructs. Optimized blue-light illumination triggered the co-localization of TALE constructs with DNMT3A-CD or TET1-CD fusion proteins at the targeted site of the Ascl1 promoter. We found that this spatiotemporal association of the fusion proteins selectively alters the methylation state and also regulates gene activity. This proof of concept developed herein holds immense promise for the ability to regulate gene activity via epigenetic modulation with spatiotemporal precision.

PMID:
28181538
PMCID:
PMC5299429
DOI:
10.1038/srep42047
[PubMed - in process]
Free PMC Article
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