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Embryonic stem cell–specific microRNAs regulate the G1-S transition and promote rapid proliferation


Dgcr8 knockout embryonic stem (ES) cells lack microprocessor activity and hence all canonical microRNAs (miRNAs). These cells proliferate slowly and accumulate in G1 phase of the cell cycle1. Here, by screening a comprehensive library of individual miRNAs in the background of the Dgcr8 knockout ES cells, we report that multiple ES cell–specific miRNAs, members of the miR-290 family, rescue the ES cell proliferation defect. Furthermore, rescued cells no longer accumulate in the G1 phase of the cell cycle. These miRNAs function by suppressing several key regulators of the G1-S transition. These results show that post-transcriptional regulation by miRNAs promotes the G1-S transition of the ES cell cycle, enabling rapid proliferation of these cells. Our screening strategy provides an alternative and powerful approach for uncovering the role of individual miRNAs in biological processes, as it overcomes the common problem of redundancy and saturation in the miRNA system.

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Figure 1: Screening for miRNAs that rescue the proliferation defects of Dgcr8 knockout (Dgcr8Δ/Δ) ES cells.
Figure 2: Rescue of proliferation and G1 accumulation defects by representative miRNAs.
Figure 3: Expression of Cdkn1a mRNA and protein upon introduction of ESCC miRNAs.
Figure 4: Luciferase reporter assay indicates that ESCC miRNAs directly interact with the 3′ UTR of Cdkn1a.
Figure 5: Additional inhibitors of the cyclin E–Cdk2 pathway are regulated by miRNAs in ES cells.

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We would like to thank A. D. Sharma and K. Okamura for their technical assistance with the immunoblot assay; M. Grskovic and C. Chaivorapol for their technical assistance with the luciferase reporter assay; M. Sachs for help in developing the overexpression construct; and M. Conti, M. Ramalho-Santos, J. Reiter and members of the Blelloch laboratory for critical reading of the manuscript. This work was supported by funds to R.B. from the US National Institutes of Health (K08 NS48118 and R01 NS057221), the Stem Cell Research Foundation and the Pew Charitable Trust. Y.W. is supported by a California Institute of Regenerative Medicine Training Grant.

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Authors and Affiliations



Y.W. performed all experiments except those shown in Figure 3a, which were done by J.E.B. S.B. developed and provided the miRNA mimic library. A.S. built mutant reporters for Figure 4. L.B. provided technical support. Y.W. and R.B. designed all experiments, analyzed data and wrote the manuscript.

Note: Supplementary information is available on the Nature Genetics website.

Corresponding author

Correspondence to Robert Blelloch.

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Competing interests

Scott Baskerville was an employee of Dharmacon/Thermo Fisher at the time of this study. He, in the form of a collaboration, supplied the microRNA mimic library used in experiments described in the manuscript.

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Supplementary Figures 1–6 and Supplementary Tables 1 and 2 (PDF 2120 kb)

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Wang, Y., Baskerville, S., Shenoy, A. et al. Embryonic stem cell–specific microRNAs regulate the G1-S transition and promote rapid proliferation. Nat Genet 40, 1478–1483 (2008).

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