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Sall4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1

Abstract

Embryonic stem (ES) cells are pluripotent cells that can self-renew or differentiate into many cell types. A unique network of transcription factors and signalling molecules are essential for maintaining this capability. Here, we report that a spalt family member, Sall4, is required for the pluripotency of ES cells. Similarly to Oct4, a reduction in Sall4 levels in mouse ES cells results in respecification, under the appropriate culture conditions, of ES cells to the trophoblast lineage. Sall4 regulates transcription of Pou5f1 which encodes Oct4. Sall4 binds to the highly conserved regulatory region of the Pou5f1 distal enhancer and activates Pou5f1 expression in vivo and in vitro. Microinjection of Sall4 small interfering (si) RNA into mouse zygotes resulted in reduction of Sall4 and Oct4 mRNAs in preimplantation embryos and significant expansion of Cdx2 expression into the inner cell mass. These results demonstrate that Sall4 is a transcriptional activator of Pou5f1 and has a critical role in the maintenance of ES cell pluripotency by modulating Oct4 expression. The data also indicates that Sall4 is important for early embryonic cell-fate decisions.

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Figure 1: Sall4 is required for ES cell self-renewal and regulates Oct4 in a dose-dependent manner.
Figure 2: Sall4 deficient ES cells differentiate and contribute to trophectoderm formation in mouse blastocyst.
Figure 3: Sall4 is able to modulate Pou5f1 transcripts in ES cells and RoSH2 cells.
Figure 4: Sall4 binds and regulates the Pou5f1 promoter.
Figure 5: Sall4 and Pou5f1 gene expression in the early mouse embryo are tightly coregulated and act in tandem with Nanog to control ES cell fate.

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Acknowledgements

This work is supported by the Agency for Science, Technology and Research (Singapore). W.L.T. is a recipient of the A*STAR graduate scholarship. The work is also partially supported by National Institutes of Health (NIH) grants to B.L. (DK04763 and AI54973), Y.M. (K08 CA097185 and P20 RR016464) and L.C. (K08 DK063220). We are grateful to colleagues in the Genome Institute of Singapore for helpful discussions and J. Buhlman for critical reading of the manuscript.

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J.Z., W.L.T. and B.L. were involved in project planning, experimental work, data analysis and manuscript preparation; G.Q.T. and H.Y.C. performed the embryo work; Q.W. performed chromatin immunoprecipitations; B.S.S. performed human ES cell culture; J.Y., Y.M. and L.C. generated knockout ES cells; and H.H.N., T.L. and P.R. were involved in project planning and manuscript preparation.

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Correspondence to Bing Lim.

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The authors declare no competing financial interests.

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Zhang, J., Tam, WL., Tong, G. et al. Sall4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1. Nat Cell Biol 8, 1114–1123 (2006). https://doi.org/10.1038/ncb1481

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