Editing DNA Methylation in the Mammalian Genome
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 inacti...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
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Elsevier
2018
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| Online Access: | http://hdl.handle.net/1721.1/116619 https://orcid.org/0000-0002-7256-3158 https://orcid.org/0000-0001-8855-8647 |
| _version_ | 1811080699501871104 |
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| author | 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. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology 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. |
| author_sort | Liu, X. Shawn |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:35:22Z |
| format | Article |
| id | mit-1721.1/116619 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T11:35:22Z |
| publishDate | 2018 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1166192022-09-27T20:33:13Z Editing DNA Methylation in the Mammalian Genome 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. Massachusetts Institute of Technology. Department of Biology Dadon, Daniel Benjamin Young, Richard A Jaenisch, Rudolf 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. National Institutes of Health (U.S.) (Grant HD045022) National Institutes of Health (U.S.) (Grant R37-CA084198) National Institutes of Health (U.S.) (Grant HG002668) National Institutes of Health (U.S.) (Grant GM114864) 2018-06-26T14:57:06Z 2018-06-26T14:57:06Z 2016-09 2016-06 2018-06-26T13:19:08Z Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/116619 Liu, X. Shawn et al. “Editing DNA Methylation in the Mammalian Genome.” Cell 167, 1 (September 2016): 233–247 © 2016 Elsevier Inc https://orcid.org/0000-0002-7256-3158 https://orcid.org/0000-0001-8855-8647 http://dx.doi.org/10.1016/J.CELL.2016.08.056 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | 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. Editing DNA Methylation in the Mammalian Genome |
| title | Editing DNA Methylation in the Mammalian Genome |
| title_full | Editing DNA Methylation in the Mammalian Genome |
| title_fullStr | Editing DNA Methylation in the Mammalian Genome |
| title_full_unstemmed | Editing DNA Methylation in the Mammalian Genome |
| title_short | Editing DNA Methylation in the Mammalian Genome |
| title_sort | editing dna methylation in the mammalian genome |
| url | http://hdl.handle.net/1721.1/116619 https://orcid.org/0000-0002-7256-3158 https://orcid.org/0000-0001-8855-8647 |
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