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dc.contributor.authorParrilla-Doblas, Jara
dc.contributor.authorAriza, Rafael R.
dc.contributor.authorRoldán-Arjona, Teresa
dc.date.accessioned2019-02-19T10:44:26Z
dc.date.available2019-02-19T10:44:26Z
dc.date.issued2017
dc.identifier.urihttp://hdl.handle.net/10396/17885
dc.description.abstractDNA methylation is a crucial epigenetic mark associated to gene silencing, and its targeted removal is amajor goal of epigenetic editing. In animal cells, DNA demethylation involves iterative 5mC oxidation byTET enzymes followed by replication-dependent dilution and/or replication-independent DNA repair of itsoxidized derivatives. In contrast, plants use specific DNA glycosylases that directly excise 5mC and initiateits substitution for unmethylated C in a base excision repair process. In this work, we have fused thecatalytic domain ofArabidopsisROS1 5mC DNA glycosylase (ROS1_CD) to the DNA binding domain ofyeast GAL4 (GBD). We show that the resultant GBD-ROS1_CD fusion protein binds specifically a GBD-targeted DNA sequencein vitro. We also found that transientin vivoexpression of GBD-ROS1_CD inhuman cells specifically reactivates transcription of a methylation-silenced reporter gene, and that suchreactivation requires both ROS1_CD catalytic activity and GBD binding capacity. Finally, we show thatreactivation induced by GBD-ROS1_CD is accompanied by decreased methylation levels at several CpGsites of the targeted promoter. All together, these results show that plant 5mC DNA glycosylases can beused for targeted active DNA demethylation in human cells.es_ES
dc.format.mimetypeapplication/pdfes_ES
dc.language.isoenges_ES
dc.publisherTaylor & Francises_ES
dc.rightshttps://creativecommons.org/licenses/by-nc-nd/4.0/es_ES
dc.sourceEpigenetics 12 (4), 296-303 (2017)es_ES
dc.subjectBase excision repaires_ES
dc.subjectDNA demethylationes_ES
dc.subjectDNA glycosylasees_ES
dc.subjectDNA methylationes_ES
dc.subjectEpigeneticeditinges_ES
dc.subject5-methylcytosinees_ES
dc.subjectTranscriptional silencinges_ES
dc.titleTargeted DNA demethylation in human cells by fusion of a plant 5-methylcytosineDNA glycosylase to a sequence-specific DNA binding domaines_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.relation.publisherversionhttp://dx.doi.org/10.1080/15592294.2017.1294306es_ES
dc.relation.projectIDGobierno de España. BFU2013-43269es_ES
dc.relation.projectIDJunta de Andalucía. P11-CVI-7576es_ES
dc.rights.accessRightsinfo:eu-repo/semantics/openAccesses_ES


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