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Updated: Sep 11, 2025
Author Spotlight: A Pipeline to Analyze Lineage-Specific Mutant Embryos at Single-Cell Resolution
Published on: June 14, 2024
RNA-Seq dataset of wild type and PRC2 mutant mouse embryonic stem cells undergoing ground state conversion
Cameron M Chapa1,2, Dustin R Fetch3, Amina A Jumamyradova1
1Department of Neuroscience, Developmental and Regenerative Biology, The University of Texas at San Antonio, San Antonio, TX 78249, USA.
Abstract:
Mouse embryonic stem cells (mESCs) represent an exceptional model for understanding how transcriptional responses are regulated by signalling pathways during development. Treatment with a cocktail of MEK and GSK3β inhibitors ("2i") induces ground state pluripotency, characterized by increased self-renewal, reduced DNA methylation, and uniformly high expression of pluripotency markers. Polycomb Repressive Complex 2 (PRC2) is a key developmental regulator controlling stem cell self-renewal and differentiation decisions, and altered expression of PRC2 target genes is a signature of 2i-mediated ground state conversion. Here, we generated a comprehensive RNA sequencing dataset from mESCs subjected to 2i conversion time-course across five time points between 0 and 96 h. We analysed two independently derived wild type lines, and two isogenic Cas9-edited lines carrying loss-of-function mutations in core PRC2 subunits Enhancer of Zeste Homolog 2 (Ezh2) or Embryonic Ectoderm Development (Eed). These data provide a resource to understand the temporal patterns of transcriptional responses to MEK and GSK3β inhibitors and explore the role of PRC2 function in regulation of pluripotency circuit.
Insights
This study details how MEK and GSK3β inhibitors (2i) induce ground state pluripotency in mouse embryonic stem cells (mESCs). It investigates the role of Polycomb Repressive Complex 2 (PRC2) in this crucial developmental process.
Area of Science:
- Stem cell biology
- Developmental biology
- Epigenetics
Background:
- Mouse embryonic stem cells (mESCs) are vital for studying developmental regulation.
- The "2i" inhibitor cocktail (MEK and GSK3β inhibitors) promotes ground state pluripotency in mESCs.
- Polycomb Repressive Complex 2 (PRC2) is critical for regulating stem cell self-renewal and differentiation.
Purpose of the Study:
- To generate a comprehensive RNA sequencing dataset of mESCs undergoing 2i-mediated ground state conversion.
- To analyze the temporal transcriptional responses to 2i treatment.
- To investigate the role of PRC2 in regulating the pluripotency circuit during 2i conversion.
Main Methods:
- Time-course RNA sequencing of mESCs treated with 2i inhibitors.
- Analysis of wild-type and PRC2-deficient (Ezh2, Eed) mESC lines.
- Bioinformatic analysis of transcriptional changes over 96 hours.
Main Results:
- Detailed temporal gene expression profiles during 2i-induced pluripotency.
- Identification of key transcriptional events and pathways regulated by 2i treatment.
- Comparative analysis highlighting PRC2's role in modulating these responses.
Conclusions:
- The generated dataset provides a valuable resource for understanding 2i-mediated pluripotency.
- PRC2 plays a significant role in the transcriptional reprogramming of mESCs.
- These findings advance our knowledge of stem cell regulation and developmental pathways.
