Cnot1, Cnot2, and Cnot3 Maintain Mouse and Human ESC Identity and Inhibit Extraembryonic Differentiation§

Authors

  • Xiaofeng Zheng,

    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    Search for more papers by this author
  • Raluca Dumitru,

    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    Search for more papers by this author
  • Brad L. Lackford,

    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    Search for more papers by this author
  • Johannes M. Freudenberg,

    1. Biostatistic Branch, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA
    Search for more papers by this author
  • Ajeet P. Singh,

    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    Search for more papers by this author
  • Trevor K. Archer,

    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    Search for more papers by this author
  • Raja Jothi,

    1. Biostatistic Branch, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA
    Search for more papers by this author
  • Guang Hu

    Corresponding author
    1. Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, RTP, North Carolina, USA
    • Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Bldg 101, Rm D-416, 111 Alexander Dr., RTP, North Carolina 27709, USA
    Search for more papers by this author
    • Telephone: 919-541-4755; Fax: 919-541-0146


  • Author contributions: G.H.: designed the research; X.Z., R.D., B.L., G.H., and A.S.: performed the research; J.F. and R.J.: performed data analysis; G.H., R.J., and T.A.: interpreted the results; G.H. and X.Z.: wrote the manuscript. R.D., B.L.L., J.M.F. contributed equally to this article.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS February 24, 2012.

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. STEM CELLS 2012;30:910–922

Ancillary