Editing DNA Methylation in the Mammalian Genome
Name
nihms813126.pdf
Size
3.29 MB
Format
Adobe PDF
Checksum (MD5)
bec50b87bde1a05b6190002d522981b2
Author(s)
Liu, X. Shawn
Wu, Hao
Ji, Xiong
Stelzer, Yonatan
Czauderna, Szymon
Shu, Jian
Dadon, Daniel Benjamin
Jaenisch, Rudolf
Wu, Xuebing, Ph. D. Massachusetts Institute of Technology
Young, Richard A.
Date Issued
September 2016
Journal
Cell
Publisher
Elsevier
Citation
Liu, X. Shawn et al. “Editing DNA Methylation in the Mammalian Genome.” Cell 167, 1 (September 2016): 233–247 © 2016 Elsevier Inc
Version
Final published version
Abstract
Mammalian DNA methylation is a critical epigenetic mechanism orchestrating gene expression networks in many biological processes. However, investigation of the functions of specific methylation events remains challenging. Here, we demonstrate that fusion of Tet1 or Dnmt3a with a catalytically inactive Cas9 (dCas9) enables targeted DNA methylation editing. Targeting of the dCas9-Tet1 or -Dnmt3a fusion protein to methylated or unmethylated promoter sequences caused activation or silencing, respectively, of an endogenous reporter. Targeted demethylation of the BDNF promoter IV or the MyoD distal enhancer by dCas9-Tet1 induced BDNF expression in post-mitotic neurons or activated MyoD facilitating reprogramming of fibroblasts into myoblasts, respectively. Targeted de novo methylation of a CTCF loop anchor site by dCas9-Dnmt3a blocked CTCF binding and interfered with DNA looping, causing altered gene expression in the neighboring loop. Finally, we show that these tools can edit DNA methylation in mice, demonstrating their wide utility for functional studies of epigenetic regulation.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.CELL.2016.08.056
