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Updated: May 24, 2026
Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors
Published on: December 3, 2017
Cnot1, Cnot2, and Cnot3 maintain mouse and human ESC identity and inhibit extraembryonic differentiation
Xiaofeng Zheng1, Raluca Dumitru, Brad L Lackford
1Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA.
Abstract:
Embryonic stem cell (ESC) identity and self-renewal is maintained by extrinsic signaling pathways and intrinsic gene regulatory networks. Here, we show that three members of the Ccr4-Not complex, Cnot1, Cnot2, and Cnot3, play critical roles in maintaining mouse and human ESC identity as a protein complex and inhibit differentiation into the extraembryonic lineages. Enriched in the inner cell mass of blastocysts, these Cnot genes are highly expressed in ESC and downregulated during differentiation. In mouse ESCs, Cnot1, Cnot2, and Cnot3 are important for maintenance in both normal conditions and the 2i/LIF medium that supports the ground state pluripotency. Genetic analysis indicated that they do not act through known self-renewal pathways or core transcription factors. Instead, they repress the expression of early trophectoderm (TE) transcription factors such as Cdx2. Importantly, these Cnot genes are also necessary for the maintenance of human ESCs, and silencing them mainly lead to TE and primitive endoderm differentiation. Together, our results indicate that Cnot1, Cnot2, and Cnot3 represent a novel component of the core self-renewal and pluripotency circuitry conserved in mouse and human ESCs.
Insights
Three Cnot genes (Cnot1, Cnot2, Cnot3) are crucial for maintaining embryonic stem cell (ESC) identity and pluripotency in mice and humans. They prevent differentiation into extraembryonic lineages by repressing key transcription factors.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Gene Regulation
Background:
- Embryonic stem cell (ESC) identity and self-renewal depend on complex signaling pathways and gene networks.
- The Ccr4-Not complex is involved in gene regulation, but its role in ESCs is not fully understood.
Purpose of the Study:
- To investigate the role of Ccr4-Not complex members (Cnot1, Cnot2, Cnot3) in maintaining mouse and human ESC identity and pluripotency.
- To determine the mechanism by which these genes influence ESC differentiation.
Main Methods:
- Analysis of Cnot gene expression in mouse and human ESCs during differentiation.
- Genetic manipulation (e.g., silencing) of Cnot genes in ESCs.
- Assessing the impact of Cnot gene manipulation on ESC identity and lineage specification.
Main Results:
- Cnot1, Cnot2, and Cnot3 are highly expressed in ESCs and downregulated upon differentiation.
- These Cnot genes are essential for maintaining pluripotency in both normal conditions and 2i/LIF medium.
- Silencing Cnot genes leads to differentiation into extraembryonic lineages, primarily trophectoderm (TE).
- Cnot genes repress the expression of TE transcription factors like Cdx2.
Conclusions:
- Cnot1, Cnot2, and Cnot3 are critical components of the core circuitry maintaining ESC self-renewal and pluripotency.
- These findings reveal a novel conserved mechanism in both mouse and human ESCs that prevents differentiation into extraembryonic lineages.
