Polycomb group RING finger protein 5 influences several developmental signaling pathways during the in vitro differentiation of mouse embryonic stem cells

• Published in: Development, growth & differentiation Vol. 62; no. 4; pp. 232 - 242
• Main Authors: Meng, Ying, Liu, Yang, Dakou, Eleni, Gutierrez, Gustavo J., Leyns, Luc
• Format: Journal Article
• Language: English
• Published: Japan Wiley Subscription Services, Inc 01.05.2020
• Subjects: Animals; Cardiomyocytes; Cell Differentiation; Cell Line; CRISPR; CRISPR-Cas Systems - genetics; CRISPR/Cas9; Ectoderm; Embryo cells; embryonic stem cells; gastrulation; Gene expression; Gene silencing; Mesenchyme; Mice; Mice, Knockout; Morphogenesis; Mouse Embryonic Stem Cells - cytology; Mouse Embryonic Stem Cells - metabolism; Notch signaling; Pcgf5; Polycomb group proteins; Polycomb-Group Proteins - deficiency; Polycomb-Group Proteins - genetics; Polycomb-Group Proteins - metabolism; Signal transduction; Signal Transduction - genetics; Stem cell transplantation; Stem cells; Wnt protein; CRISPR/Cas9; Notch signaling; Pcgf5; embryonic stem cells; gastrulation
• ISSN: 0012-1592; 1440-169X
• DOI: 10.1111/dgd.12659

Polycomb group (PcG) RING finger protein 5 (PCGF5) is a core component of the so‐called Polycomb repressive complex 1.5 (PRC1.5), which is involved in epigenetic transcriptional repression. To explore the developmental function of Pcgf5, we generated Pcgf5 knockout (Pcgf5−/−) mouse embryonic stem cell (mESC) lines with the help of CRISPR/Cas9 technology. We subjected the Pcgf5−/− and wild‐type (WT) mESCs to a differentiation protocol toward mesodermal‐cardiac cell types as aggregated embryoid bodies (EBs) and we found that knockout of Pcgf5 delayed the generation of the three germ layers, especially the ectoderm. Further, disruption of Pcgf5 impacted the epithelial‐mesenchymal transition during EB morphogenesis and differentially affected the gene expression of essential developmental signaling pathways such as Nodal and Wnt. Finally, we also unveiled that loss of Pcgf5 induced the repression of genes involved in the Notch pathway, which may explain the enhancement of cardiomyocyte maturation and the dampening of ectodermal‐neural differentiation observed in the Pcgf5−/− EBs. Pcgf5 knockout delays the generation of the three germ layers during differentiation, especially the ectoderm. The disruption of Pcgf5 impacts the epithelial‐mesenchymal transition and affects essential developmental signaling pathways, such as Nodal or Wnt. The loss of Pcgf5 represses the Notch pathway, which may explain the dampened ectodermal‐neural differentiation.