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A core Klf circuitry regulates self-renewal of embryonic stem cells

Abstract

Embryonic stem (ES) cells are unique in their ability to self-renew indefinitely and maintain pluripotency. These properties require transcription factors that specify the gene expression programme of ES cells. It has been possible to reverse the highly differentiated state of somatic cells back to a pluripotent state with a combination of four transcription factors: Klf4 is one of the reprogramming factors required, in conjunction with Oct4, Sox2 and c-Myc. Maintenance of self-renewal and pluripotency of ES cells requires Oct4, Sox2 and c-Myc, but Klf4 is dispensable. Here, we show that Krüppel-like factors are required for the self-renewal of ES cells. Simultaneous depletion of Klf2, Klf4 and Klf5 lead to ES cell differentiation. Chromatin immunoprecipitation coupled to microarray assay reveals that these Klf proteins share many common targets of Nanog, suggesting a close functional relationship between these factors. Expression analysis after triple RNA interference (RNAi) of the Klfs shows that they regulate key pluripotency genes, such as Nanog. Taken together, our study provides new insight into how the core Klf circuitry integrates into the Nanog transcriptional network to specify gene expression that is unique to ES cells.

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Figure 1: Klf4, Klf2 and Klf5 are required for the maintenance of ES cells
Figure 2: Identification of in vivo binding sites of Klf4, Klf2 and Klf5 by ChIP-on-chip.
Figure 3: Enhancer activity of Klf bound regions.
Figure 4: Global gene expression changes after triple knockdown of Klf proteins.
Figure 5: Integration of the core Klf circuitry with the Nanog transcriptional regulatory network in ES cells (a) Schematic representation of a model for the transcriptional regulatory network targeted by Klf2, Klf4, Klf5 and Nanog.

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Acknowledgements

We are grateful to the Biomedical Research Council (BMRC) and Agency for Science, Technology and Research (A*STAR) for funding. J.J is supported by the Singapore Millennium Foundation graduate scholarship and the NUS graduate scholarship. Y.-H.L. and C.-A.L. are supported by the A*STAR graduate scholarships. We are grateful to T. Lufkin for D3 ES cells and K.Kuay and D.-X. Lin for technical support. We thank T. Huber and E. Cheung for critical comments on the manuscript.

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Correspondence to Huck-Hui Ng.

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Jiang, J., Chan, YS., Loh, YH. et al. A core Klf circuitry regulates self-renewal of embryonic stem cells. Nat Cell Biol 10, 353–360 (2008). https://doi.org/10.1038/ncb1698

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