Characterization of Mammalian Regulatory Complexes at Single-Locus Resolution Using TINC

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dc.contributor.authorKnaupp, A.S.
dc.contributor.authorSchittenhelm, R.B.
dc.contributor.authorPolo, J.M.
dc.contributor.editorHorsfield, J.
dc.contributor.editorMarsman, J.
dc.date.issued2022
dc.descriptionBook series - Print ISSN: 1064-3745 ; Electronic ISSN: 1940-6029
dc.description.abstractIn mammalian cells, multiprotein complexes form at specific genomic regulatory elements (REs) to control gene expression, which in turn is ultimately responsible for cellular identity. Consequently, insight into the molecular composition of these regulatory complexes is of major importance for our understanding of any physiological or pathological cellular state or transition. However, it remains extremely difficult to identify the protein complex(es) assembled at a specific RE in the mammalian genome using conventional approaches. We therefore developed a novel single locus isolation technique based on Transcription Activator-Like Effector (TALE) proteins termed TALE-mediated isolation of nuclear chromatin (TINC). When coupled with high-resolution mass spectrometry, TINC enables the identification and characterization of protein complexes formed at any RE of interest. Using the Nanog promoter in mouse embryonic stem cells as proof of concept, this chapter describes in detail the novel TINC methodology as well as subsequent mass spectrometric considerations.
dc.description.statementofresponsibilityAnja S. Knaupp, Ralf B. Schittenhelm, and Jose M. Polo
dc.identifier.citationChromatin: Methods and Protocols, 2022 / Horsfield, J., Marsman, J. (ed./s), vol.2458, Ch.10, pp.175-193
dc.identifier.doi10.1007/978-1-0716-2140-0_10
dc.identifier.isbn9781071621394
dc.identifier.orcidPolo, J.M. [0000-0002-2531-778X]
dc.identifier.urihttps://hdl.handle.net/2440/136966
dc.language.isoen
dc.publisherHumana
dc.publisher.placeNew York, NY
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1069830
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1036587
dc.relation.granthttp://purl.org/au-research/grants/arc/FT180100674
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1092280
dc.relation.granthttp://purl.org/au-research/grants/arc/DP210104029
dc.relation.ispartofseriesMethods in Molecular Biology; 2458
dc.rights© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2022
dc.source.urihttps://link.springer.com/book/10.1007/978-1-0716-2140-0
dc.subjectSingle-locus pulldown
dc.subjectEpigenetics
dc.subjectProteomics
dc.subjectRegulatory complex
dc.subjectTranscriptional regulation
dc.subjectAffinity purification
dc.subjectTranscription activator-like effector proteins
dc.subjectChromatin immuno-precipitation
dc.subjectMass spectrometry
dc.subject.meshAnimals
dc.subject.meshMammals
dc.subject.meshMice
dc.subject.meshMultiprotein Complexes
dc.subject.meshGenomics
dc.subject.meshMass Spectrometry
dc.subject.meshPromoter Regions, Genetic
dc.titleCharacterization of Mammalian Regulatory Complexes at Single-Locus Resolution Using TINC
dc.typeBook chapter
pubs.publication-statusPublished

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