Myc-regulated microRNAs attenuate embryonic stem cell differentiation

Authors

  • Chin-Hsing Lin,

    1. Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
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  • Aimee L Jackson,

    1. Department of Biology, Rosetta Inpharmatics LLC, Seattle, WA, USA
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    • Present address: Regulus Therapeutics, 1896 Rutherford Rd, Carlsbad, CA 92008, USA
  • Jie Guo,

    1. Department of Biology, Rosetta Inpharmatics LLC, Seattle, WA, USA
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  • Peter S Linsley,

    1. Department of Biology, Rosetta Inpharmatics LLC, Seattle, WA, USA
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    • Present address: Regulus Therapeutics, 1896 Rutherford Rd, Carlsbad, CA 92008, USA
  • Robert N Eisenman

    Corresponding author
    1. Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
    • Corresponding author. Division of Basic Sciences, Fred Hutchinson Cancer Research Center A2M-025, 1100 Fairview Avenue N., POB 19024, Seattle, WA 98109-4417, USA. Tel.: +1 206 667 4445; Fax: +1 206 667 6522; E-mail: eisenman@fhcrc.org

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Abstract

Myc proteins are known to have an important function in stem cell maintenance. As Myc has been shown earlier to regulate microRNAs (miRNAs) involved in proliferation, we sought to determine whether c-Myc also affects embryonic stem (ES) cell maintenance and differentiation through miRNAs. Using a quantitative primer-extension PCR assay we identified miRNAs, including, miR-141, miR-200, and miR-429 whose expression is regulated by c-Myc in ES cells, but not in the differentiated and tumourigenic derivatives of ES cells. Chromatin immunoprecipitation analyses indicate that in ES cells c-Myc binds proximal to genomic regions encoding the induced miRNAs. We used expression profiling and seed homology to identify genes specifically downregulated both by these miRNAs and by c-Myc. We further show that the introduction of c-Myc-induced miRNAs into murine ES cells significantly attenuates the downregulation of pluripotency markers on induction of differentiation after withdrawal of the ES cell maintenance factor LIF. In contrast, knockdown of the endogenous miRNAs accelerate differentiation. Our data show that in ES cells c-Myc acts, in part, through a subset of miRNAs to attenuate differentiation.

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