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  Genome-wide tracking of dCas9-methyltransferase footprints

Galonska, C., Charlton, J., Mattei, A. L., Donaghey, J., Clement, K., Gu, H., et al. (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nature Communications, 9: 9:597. doi:10.1038/s41467-017-02708-5.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0000-7667-8 Version Permalink: http://hdl.handle.net/21.11116/0000-0000-769F-9
Genre: Journal Article

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 Creators:
Galonska, Christina1, Author              
Charlton, Jocelyn1, 2, Author              
Mattei, Alexandra L.1, 2, 3, Author              
Donaghey, Julie 2, 3, Author
Clement, Kendell 2, 3, Author
Gu, Hongcang3, Author
Mohammad, Arman W. 3, Author
Stamenova, Elena K. 3, Author
Cacchiarelli, Davide 3, Author
Klages, Sven4, Author              
Timmermann, Bernd4, Author              
Cantz, Tobias , Author
Schöler, Hans R. , Author
Gnirke, Andreas 3, Author
Ziller, Michael J. , Author
Meissner, Alexander1, 2, 3, Author              
Affiliations:
1Dept. of Genome Regulation (Head: Alexander Meissner), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_2379694              
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA, ou_persistent22              
3Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA, ou_persistent22              
4Sequencing (Head: Bernd Timmermann), Scientific Service (Head: Christoph Krukenkamp), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479670              

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 Abstract: In normal mammalian development cytosine methylation is essential and is directed to specific regions of the genome. Despite notable advances through mapping its genome-wide distribution, studying the direct contribution of DNA methylation to gene and genome regulation has been limited by the lack of tools for its precise manipulation. Thus, combining the targeting capability of the CRISPR–Cas9 system with an epigenetic modifier has attracted interest in the scientific community. In contrast to profiling the genome-wide cleavage of a nuclease competent Cas9, tracing the global activity of a dead Cas9 (dCas9) methyltransferase fusion protein is challenging within a highly methylated genome. Here, we report the generation and use of an engineered, methylation depleted but maintenance competent mouse ES cell line and find surprisingly ubiquitous nuclear activity of dCas9-methyltransferases. Subsequent experiments in human somatic cells refine these observations and point to an important difference between genetic and epigenetic editing tools that require unique experimental considerations.

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Language(s): eng - English
 Dates: 2017-12-202018-02-09
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: DOI: 10.1038/s41467-017-02708-5
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Title: Nature Communications
  Abbreviation : Nat. Commun.
Source Genre: Journal
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Publ. Info: London : Nature Publishing Group
Pages: - Volume / Issue: 9 Sequence Number: 9:597 Start / End Page: - Identifier: ISSN: 2041-1723
CoNE: /journals/resource/2041-1723