Epigenetic Editing of Ascl1 Gene in Neural Stem Cells by Optogenetics

Chiao Ling Lo, Samrat Roy Choudhury, Joseph Irudayaraj, Feng C. Zhou

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article number42047
JournalScientific reports
Volume7
DOIs
StatePublished - Feb 9 2017
Externally publishedYes

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Optogenetics
Neural Stem Cells
Epigenomics
Catalytic Domain
Methylation
Genetic Epigenesis
Catalytic DNA
Genes
Dioxygenases
Proteins
Methyltransferases
Spinal Ganglia
Gene Editing
Lighting
Genetic Promoter Regions
Histones
Light
DNA
Enzymes

ASJC Scopus subject areas

  • General

Cite this

Epigenetic Editing of Ascl1 Gene in Neural Stem Cells by Optogenetics. / Lo, Chiao Ling; Choudhury, Samrat Roy; Irudayaraj, Joseph; Zhou, Feng C.

In: Scientific reports, Vol. 7, 42047, 09.02.2017.

Research output: Contribution to journalArticle

Lo, Chiao Ling ; Choudhury, Samrat Roy ; Irudayaraj, Joseph ; Zhou, Feng C. / Epigenetic Editing of Ascl1 Gene in Neural Stem Cells by Optogenetics. In: Scientific reports. 2017 ; Vol. 7.
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