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PXD021089

PXD021089 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitlegreenCUT&RUN: a rapid and efficient genome profiling approach for transcription and chromatin regulatory proteins
DescriptionChromatin modifiers, transcription factors and cofactors regulate gene expression through engagement with chromatin and DNA, which can be detected by a variety of genomic techniques. Here, we describe a novel method ‘greenCUT&RUN’ for genome-wide profiling of transcription regulatory factors, which has higher sensitivity, resolution, accuracy and reproducibility than existing methods, whilst assuring specificity as assessed for the gene specific transcription factors NFY and c-FOS, and for the basal transcription factor TBP. Our strategy begins with tagging the protein of interest with GFP and utilizes a GFP-specific nanobody fused to MNase to profile genome-wide binding events. Since the nanobody targets GFP, the greenCUT&RUN approach eliminates dependency on protein-specific antibodies, which makes it widely applicable. To examine, if GFP-tagged proteins form complex with known interactors and GFP did not interfere in the formation of protein complex, mass spectrometry was performed.
HostingRepositoryPRIDE
AnnounceDate2021-01-13
AnnouncementXMLSubmission_2021-01-12_23:39:01.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterMarc Timmers
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentOrbitrap Fusion Lumos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-08-24 09:06:20ID requested
12021-01-12 23:39:01announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: NFYA, FOS, HeLa, GFP-tagged proteins, Affinity purification, Nanobody, Enhancer
Contact List
H. Th. Marc Timmers
contact affiliation1. German Cancer Consortium (DKTK) partner site Freiburg, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany 2. Department of Urology, Medical Center-University of Freiburg, 79106 Freiburg, Germany
contact emailm.timmers@dkfz-heidelberg.de
lab head
Marc Timmers
contact affiliationGerman Cancer Consortium (DKTK), partner site Freiburg German Cancer Research Center (DKFZ) Foundation under Public Law c/o Department of Urology Zentrale Klinische Forschung (ZKF) Medical Center-University of Freiburg Breisacher Str. 66, 79106 Freiburg, Germany
contact emailm.timmers@dkfz-heidelberg.de
dataset submitter
Full Dataset Link List
Dataset FTP location
PRIDE project URI
Repository Record List
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