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dc.contributor.authorCheetham, Seth W
dc.contributor.authorGruhn, Wolfram H
dc.contributor.authorvan den Ameele, Jelle
dc.contributor.authorKrautz, Robert
dc.contributor.authorSouthall, Tony D
dc.contributor.authorKobayashi, Toshihiro
dc.contributor.authorSurani, M Azim
dc.contributor.authorBrand, Andrea H
dc.date.accessioned2018-10-03T04:45:49Z
dc.date.available2018-10-03T04:45:49Z
dc.date.issued2018-10-17
dc.identifier.issn0950-1991
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/283107
dc.description.abstractThe precise control of gene expression by transcription factor networks is crucial to organismal development. The predominant approach for mapping transcription factor-chromatin interactions has been chromatin immunoprecipitation (ChIP). However, ChIP requires a large number of homogeneous cells and antisera with high specificity. A second approach, DamID, has the drawback that high levels of Dam methylase are toxic. Here, we modify our targeted DamID approach (TaDa) to enable cell type-specific expression in mammalian systems, generating an inducible system (mammalian TaDa or MaTaDa) to identify genome-wide protein/DNA interactions in 100 to 1000 times fewer cells than ChIP-based approaches. We mapped the binding sites of two key pluripotency factors, OCT4 and PRDM14, in mouse embryonic stem cells, epiblast-like cells and primordial germ cell-like cells (PGCLCs). PGCLCs are an important system for elucidating primordial germ cell development in mice. We monitored PRDM14 binding during the specification of PGCLCs, identifying direct targets of PRDM14 that are key to understanding its crucial role in PGCLC development. We show that MaTaDa is a sensitive and accurate method for assessing cell type-specific transcription factor binding in limited numbers of cells.
dc.format.mediumElectronic
dc.languageeng
dc.publisherThe Company of Biologists
dc.subjectGerm Cells
dc.subjectChromatin
dc.subjectPluripotent Stem Cells
dc.subjectAnimals
dc.subjectMice
dc.subjectRNA-Binding Proteins
dc.subjectDNA-Binding Proteins
dc.subjectTranscription Factors
dc.subjectDNA Methylation
dc.subjectBinding Sites
dc.subjectProtein Binding
dc.subjectGenome
dc.subjectMouse Embryonic Stem Cells
dc.titleTargeted DamID reveals differential binding of mammalian pluripotency factors.
dc.typeArticle
prism.issueIdentifier20
prism.publicationDate2018
prism.publicationNameDevelopment
prism.volume145
dc.identifier.doi10.17863/CAM.30468
dcterms.dateAccepted2018-08-23
rioxxterms.versionofrecord10.1242/dev.170209
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2018-10-17
dc.contributor.orcidCheetham, Seth W [0000-0001-6428-3175]
dc.contributor.orcidvan den Ameele, Jelle [0000-0002-2744-0810]
dc.contributor.orcidSurani, M Azim [0000-0002-8640-4318]
dc.contributor.orcidBrand, Andrea H [0000-0002-2089-6954]
dc.identifier.eissn1477-9129
rioxxterms.typeJournal Article/Review
pubs.funder-project-idWellcome Trust (092545/Z/10/Z)
pubs.funder-project-idWellcome Trust (103792/Z/14/Z)
pubs.funder-project-idWellcome Trust (105839/Z/14/Z)
pubs.funder-project-idRoyal Society (RP150061)
pubs.funder-project-idWellcome Trust (096738/Z/11/Z)
pubs.funder-project-idWellcome Trust (092096/Z/10/Z)
pubs.funder-project-idCancer Research Uk (None)
cam.issuedOnline2018-10-17
rioxxterms.freetoread.startdate2019-09-05


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