Profiling of pluripotency factors in individual stem cells and early embryos [preprint]
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyDepartment of Molecular, Cell, and Cancer Biology
Document Type
PreprintPublication Date
2018-03-21Keywords
pluripotencystem cells
embryos
genomics
Cell Biology
Cells
Developmental Biology
Embryonic Structures
Genomics
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Show full item recordAbstract
Major cell fate decisions are governed by sequence-specific transcription factors (TFs) that act in small cell populations within developing embryos. To understand how TFs regulate cell fate it is important to identify their genomic binding sites in these populations. However, current methods cannot profile TFs genome-wide at or near the single cell level. Here we adapt the CUT&RUN method to profile chromatin proteins in low cell numbers, mapping TF-DNA interactions in single cells and individual pre-implantation embryos for the first time. Using this method, we demonstrate that the pluripotency TF NANOG is significantly more dependent on the SWI/SNF family ATPase BRG1 for association with its genomic targets in vivo than in cultured cells, a finding that could not have been made using traditional approaches. Ultra-low input CUT&RUN (uliCUT&RUN) enables interrogation of TF binding from low cell numbers, with broad applicability to rare cell populations of importance in development or disease.Source
bioRxiv 286351; doi: https://doi.org/10.1101/286351. Link to preprint on bioRxiv service.
DOI
10.1101/286351Permanent Link to this Item
http://hdl.handle.net/20.500.14038/29277Rights
The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.Distribution License
http://creativecommons.org/licenses/by-nc-nd/4.0/ae974a485f413a2113503eed53cd6c53
10.1101/286351
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Except where otherwise noted, this item's license is described as The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.