Targeted DamID reveals differential binding of mammalian pluripotency factors
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
The 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.
Date Issued
2018-10-15
Date Acceptance
2018-08-23
Citation
Development, 2018, 145 (20)
ISSN
0950-1991
Publisher
Company of Biologists Ltd
Journal / Book Title
Development
Volume
145
Issue
20
Copyright Statement
© 2018 The Author(s). Published by The Company of Biologists Ltd. http://www.biologists.com/user-licence-1-1/
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000449152600014&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Life Sciences & Biomedicine
Developmental Biology
ChIP-seq
Embryonic stem cells
Oct4
Prdm14
Primordial germ cells
Targeted DamID
PROTEIN-DNA INTERACTIONS
GERM-CELL SPECIFICATION
EMBRYONIC STEM-CELLS
NAIVE PLURIPOTENCY
GENE-EXPRESSION
READ ALIGNMENT
IN-VITRO
MOUSE
PRDM14
INDUCTION
Publication Status
Published
Article Number
dev170209
Date Publish Online
2018-10-17