WDR5, BRCA1, and BARD1 Co-regulate the DNA Damage Response and Modulate the Mesenchymal-to-Epithelial Transition during Early Reprogramming.
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ICR Authors
Authors
Peñalosa-Ruiz, G
Bousgouni, V
Gerlach, JP
Waarlo, S
van de Ven, JV
Veenstra, TE
Silva, JCR
van Heeringen, SJ
Bakal, C
Mulder, KW
Veenstra, GJC
Bousgouni, V
Gerlach, JP
Waarlo, S
van de Ven, JV
Veenstra, TE
Silva, JCR
van Heeringen, SJ
Bakal, C
Mulder, KW
Veenstra, GJC
Document Type
Journal Article
Date
2019-04-09
Date Accepted
2019-02-14
Date Available
Abstract
Differentiated cells are epigenetically stable, but can be reprogrammed to pluripotency by expression of the OSKM transcription factors. Despite significant effort, relatively little is known about the cellular requirements for reprogramming and how they affect the properties of induced pluripotent stem cells. We have performed high-content screening with small interfering RNAs targeting 300 chromatin-associated factors and extracted colony-level quantitative features. This revealed five morphological phenotypes in early reprogramming, including one displaying large round colonies exhibiting an early block of reprogramming. Using RNA sequencing, we identified transcriptional changes associated with these phenotypes. Furthermore, double knockdown epistasis experiments revealed that BRCA1, BARD1, and WDR5 functionally interact and are required for the DNA damage response. In addition, the mesenchymal-to-epithelial transition is affected in Brca1, Bard1, and Wdr5 knockdowns. Our data provide a resource of chromatin-associated factors in early reprogramming and underline colony morphology as an important high-dimensional readout for reprogramming quality.
Citation
Stem cell reports, 2019, 12 (4), pp. 743 - 756
Source Title
Publisher
CELL PRESS
ISSN
2213-6711
eISSN
2213-6711
Collections
Research Team
Dynamical Cell Systems