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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Keller, G. Embryonic stem cell differentiation: emergence of a new era in biology and medicine. Genes Dev. 19, 1129–1155 (2005).
Surani, M. A., Hayashi, K. & Hajkova, P. Genetic and epigenetic regulators of pluripotency. Cell 128, 747–762 (2007).
Park, I. H. & Daley, G. Q. Debugging cellular reprogramming. Nature Cell. Biol. 9, 871–873 (2007).
Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006).
Okita, K., Ichisaka, T. & Yamanaka, S. Generation of germline-competent induced pluripotent stem cells. Nature 448, 313–317 (2007).
Wernig, M. et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 448, 318–324 (2007).
Maherali, N. et al. Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell 1, 55–71 (2007).
Niwa, H., Miyazaki, J. & Smith, A. G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nature Genet. 24, 372–376 (2000).
Ivanova, N. et al. Dissecting self-renewal in stem cells with RNA interference. Nature 442, 533–538 (2006).
Loh, Y. H. et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nature Genet. 38, 431–440 (2006).
Masui, S. et al. Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells. Nature Cell. Biol. 9, 625–635 (2007).
Cartwright, P. et al. LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism. Development 132, 885–896 (2005).
McConnell, B. B., Ghaleb, A. M., Nandan, M. O. & Yang, V. W. The diverse functions of Kruppel-like factors 4 and 5 in epithelial biology and pathobiology. Bioessays 29, 549–557 (2007).
Nakatake, Y. et al. Klf4 cooperates with Oct3/4 and Sox2 to activate the Lefty1 core promoter in embryonic stem cells. Mol. Cell. Biol. 26, 7772–7782 (2006).
Matoba, R. et al. Dissecting Oct3/4-regulated gene networks in embryonic stem cells by expression profiling. PLoS 1, e26 (2006).
Zhang, J. et al. Sall4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1. Nature Cell. Biol. 8, 1114–1123 (2006).
Nichols, J. et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95, 379–391 (1998).
Avilion, A. A. et al. Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev. 17, 126–140 (2003).
Anisimov, S. V. et al. SAGE identification of gene transcripts with profiles unique to pluripotent mouse R1 embryonic stem cells. Genomics 79, 169–176 (2002).
Hong, Y., Cervantes, R. B., Tichy, E., Tischfield, J. A. & Stambrook, P. J. Protecting genomic integrity in somatic cells and embryonic stem cells. Mutat. Res. 614, 48–55 (2007).
Mitsui, K. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642 (2003).
Chambers, I. et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655 (2003).
Zhang, W. et al. The gut-enriched Kruppel-like factor (Kruppel-like factor 4) mediates the transactivating effect of p53 on the p21WAF1/Cip1 promoter. J. Biol. Chem. 275, 18391–18398 (2000).
Brons, I. G. et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 448, 191–195 (2007).
Tesar, P. J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196–199 (2007).
Li, Y. et al. Murine embryonic stem cell differentiation is promoted by SOCS-3 and inhibited by the zinc finger transcription factor Klf4. Blood 105, 635–637 (2005).
Rowland, B. D., Bernards, R. & Peeper, D. S. The KLF4 tumour suppressor is a transcriptional repressor of p53 that acts as a context-dependent oncogene. Nature Cell. Biol. 7, 1074–1082 (2005).
Wani, M. A., Wert, S. E. & Lingrel, J. B. Lung Kruppel-like factor, a zinc finger transcription factor, is essential for normal lung development. J. Biol. Chem. 274, 21180–21185 (1999).
Kuo, C. T. et al. The LKLF transcription factor is required for normal tunica media formation and blood vessel stabilization during murine embryogenesis. Genes Dev. 11, 2996–3006 (1997).
Carlson, C. M. et al. Kruppel-like factor 2 regulates thymocyte and T-cell migration. Nature 442, 299–302 (2006).
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.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Information
Supplementary figures S1, S2, S3, S4, S5, S6 (PDF 1400 kb)
Supplementary Information
Supplementary Table 1 (XLS 74 kb)
Supplementary Information
Supplementary Table 2 (XLS 556 kb)
Supplementary Information
Supplementary Table 3 (XLS 1068 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1698
This article is cited by
-
G-quadruplex DNA structures in human stem cells and differentiation
Nature Communications (2022)
-
KLF17 promotes human naive pluripotency through repressing MAPK3 and ZIC2
Science China Life Sciences (2022)
-
PRMT1 and PRMT5: on the road of homologous recombination and non-homologous end joining
Genome Instability & Disease (2022)
-
Advances in RNA Viral Vector Technology to Reprogram Somatic Cells: The Paramyxovirus Wave
Molecular Diagnosis & Therapy (2022)
-
Opposing transcriptional programs of KLF5 and AR emerge during therapy for advanced prostate cancer
Nature Communications (2021)