Abstract
Arising from: S. K. Singh et al. Nature 453, 223–227 (2008)10.1038/nature06863; Singh et al. reply
The contribution of REST to embryonic stem (ES) cell pluripotency has been uncertain. Two years ago, Singh et al.1 claimed that Rest+/− and REST knock-down ES cells expressed reduced levels of pluripotency markers1, in contrast to a prior2 and subsequent reports3,4,5,6. To understand the basis of this difference, we analysed the YHC334 (YHC) and RRC160 (RRC) gene-trap ES cell lines used by Singh et al.1, obtained directly from BayGenomics. Both REST mutant lines generated REST–βGeo fusion proteins, but expressed pluripotency genes at levels similar to appropriately matched parental wild ES cells, consistent with expression being REST–independent.
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References
Singh, S. K., Kagalwala, M., Parker-Thornburg, J., Adams, H. & Majumder, S. REST maintains self-renewal and pluripotency of embryonic stem cells. Nature 453, 223–227 (2008)
Loh, Y. H. et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nature Genet. 38, 431–440 (2006)
Buckley, N., Johnson, R., Sun, Y. M. & Stanton, L. Is REST a regulator of pluripotency? Nature 457, E5–E6 (2009)
Jørgensen, H. F., Chen, Z. F., Merkenschlager, M. & Fisher, A. G. Is REST required for ESC pluripotency? Nature 457, E4–E5 (2009)
Jørgensen, H. F. et al. REST selectively represses a subset of RE1-containing neuronal genes in mouse embryonic stem cells. Development 136, 715–721 (2009)
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Roopra, A. et al. Transcriptional repression by neuron-restrictive silencer factor is mediated via the Sin3-histone deacetylase complex. Mol. Cell. Biol. 20, 2147–2157 (2000)
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H.F.J. performed the experiments. H.F.J. and A.G.F. conceived the analysis and wrote the manuscript.
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Jørgensen, H., Fisher, A. Can controversies be put to REST?. Nature 467, E3–E4 (2010). https://doi.org/10.1038/nature09305
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DOI: https://doi.org/10.1038/nature09305
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