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Opposing microRNA families regulate self-renewal in mouse embryonic stem cells

Nature volume 463, pages 621–626 (2010)Cite this article

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A Corrigendum to this article was published on 04 March 2010

Abstract

When embryonic stem cells (ESCs) differentiate, they must both silence the ESC self-renewal program and activate new tissue-specific programs. In the absence of DGCR8 (Dgcr8-/-), a protein required for microRNA (miRNA) biogenesis, mouse ESCs are unable to silence self-renewal. Here we show that the introduction of let-7 miRNAs—a family of miRNAs highly expressed in somatic cells—can suppress self-renewal in Dgcr8-/- but not wild-type ESCs. Introduction of ESC cell cycle regulating (ESCC) miRNAs into the Dgcr8-/- ESCs blocks the capacity of let-7 to suppress self-renewal. Profiling and bioinformatic analyses show that let-7 inhibits whereas ESCC miRNAs indirectly activate numerous self-renewal genes. Furthermore, inhibition of the let-7 family promotes de-differentiation of somatic cells to induced pluripotent stem cells. Together, these findings show how the ESCC and let-7 miRNAs act through common pathways to alternatively stabilize the self-renewing versus differentiated cell fates.

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Figure 1: The let-7 and ESCC miRNA families have opposing roles in regulating ESC self-renewal.
Figure 2: The let-7 and ESCC miRNAs suppress hundreds of transcripts by binding their ORF and/or 3′UTR.
Figure 3: Enrichment/depletion of transcription-factor-bound genes among miRNA-regulated transcripts.
Figure 4: Let-7c and miR-294 regulate Lin28, Sall4, c-Myc and N-Myc.
Figure 5: Inhibition of let-7 miRNAs promotes reprogramming to induced pluripotency.

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Primary accessions

Gene Expression Omnibus

Data deposits

Our microarray profiling data can be found at the Gene Expression Omnibus (GEO) database, under accession GSE18840.

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Acknowledgements

We would like to thank M. Ramalho-Santos, S. Fisher, M. Conti and members of the Blelloch laboratory for critical reading of the manuscript. We would like to thank A. Amiet for let-7 and control inhibitors, M. Bishop for c-Myc floxed mice and A. Shenoy for ESC-derived neural progenitor cells cultures. Furthermore, we would like to acknowledge A. Olshen and R. Roy for their helpful advice concerning our statistical methods. This work was supported by funds to R.B. from the National Institutes of Health (NIH; K08 NS48118 and R01 NS057221), California Institute of Regenerative Medicine (CIRM; Seed Grant RS1-00161, New Faculty Award RN2-00906), the American Health Assistance Foundation (formerly Stem Cell Research Foundation), and the Pew Charitable Trust. C.M. and R.L.J. are supported by the National Science Foundation (NSF) graduate research fellowships.

Author Contributions C.M. contributed to Figs 1–4, 5b and Supplementary Figs 1–13 and 18. R.L.J. contributed to Fig. 5a and Supplementary Figs 13–17. C.M., R.L.J. and R.B. conceived the experiments, analysed the data, and wrote the manuscript.

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Authors and Affiliations

  1. The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, Program in Biomedical Sciences,,

    Collin Melton, Robert L. Judson & Robert Blelloch

  2. Department of Urology, University of California San Francisco, San Francisco, California 94143, USA,

    Collin Melton, Robert L. Judson & Robert Blelloch

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  1. Collin Melton
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Correspondence to Robert Blelloch.

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Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-18 with Legends. (PDF 1424 kb)

Supplementary Table 1

This table contains microarray data with seed match information. (XLS 9141 kb)

Supplementary Table 2

This table contains pluripotency-associated genes and data used in Fig. S7. (XLS 19 kb)

Supplementary Table 3

This table contains microarray and seed match information for transcripts both up-regulated by miR-294 and down-regulated by let-7c. (XLS 25 kb)

Supplementary Table 4

This table contains qPCR primers. (XLS 21 kb)

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Melton, C., Judson, R. & Blelloch, R. Opposing microRNA families regulate self-renewal in mouse embryonic stem cells. Nature 463, 621–626 (2010). https://doi.org/10.1038/nature08725

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