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Nat Commun. 2017 Jul 11;8:16026. doi: 10.1038/ncomms16026.

Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein.

Lei Y1,2, Zhang X1,2, Su J3, Jeong M1,2, Gundry MC1,2, Huang YH2,4, Zhou Y5, Li W2, Goodell MA1,2,3.

Author information

1
Department of Molecular and Human Genetics, Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas 77030, USA.
2
Stem Cells and Regenerative Medicine Center, Baylor College of Medicine, Houston, Texas 77030, USA.
3
Dan L. Duncan Cancer Center and Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
4
Department of Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
5
Center for Translational Cancer Research, Institute of Biosciences and Technology, Department of Medical Physiology, College of Medicine, Texas A&M University, Houston, Texas 77030, USA.

Abstract

Comprehensive studies have shown that DNA methylation plays vital roles in both loss of pluripotency and governance of the transcriptome during embryogenesis and subsequent developmental processes. Aberrant DNA methylation patterns have been widely observed in tumorigenesis, ageing and neurodegenerative diseases, highlighting the importance of a systematic understanding of DNA methylation and the dynamic changes of methylomes during disease onset and progression. Here we describe a facile and convenient approach for efficient targeted DNA methylation by fusing inactive Cas9 (dCas9) with an engineered prokaryotic DNA methyltransferase MQ1. Our study presents a rapid and efficient strategy to achieve locus-specific cytosine modifications in the genome without obvious impact on global methylation in 24 h. Finally, we demonstrate our tool can induce targeted CpG methylation in mice by zygote microinjection, thereby demonstrating its potential utility in early development.

PMID:
28695892
PMCID:
PMC5508226
DOI:
10.1038/ncomms16026
[Indexed for MEDLINE]
Free PMC Article

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