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Editing DNA Methylation in the Mammalian Genome.

Cell | Sep 22, 2016

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.

Pubmed ID: 27662091 RIS Download

Mesh terms: Animals | Bacterial Proteins | Brain-Derived Neurotrophic Factor | Cell Line | Clustered Regularly Interspaced Short Palindromic Repeats | CpG Islands | DNA (Cytosine-5-)-Methyltransferase | DNA Methylation | DNA-Binding Proteins | Endonucleases | Enhancer Elements, Genetic | Epigenesis, Genetic | Gene Editing | Genome | Mice | MyoD Protein | Neurons | Promoter Regions, Genetic | Protein Binding | Proto-Oncogene Proteins | Recombinant Fusion Proteins | Repressor Proteins

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Associated grants

  • Agency: NCI NIH HHS, Id: R37 CA084198
  • Agency: NIGMS NIH HHS, Id: T32 GM007287
  • Agency: NCI NIH HHS, Id: P30 CA014051
  • Agency: NIGMS NIH HHS, Id: R01 GM114864
  • Agency: NINDS NIH HHS, Id: R01 NS088538
  • Agency: NICHD NIH HHS, Id: R37 HD045022
  • Agency: NICHD NIH HHS, Id: R01 HD045022
  • Agency: NHGRI NIH HHS, Id: R01 HG002668

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