1. TCF/LEF regulation of the topologically associated domain ADI promotes mESCs to exit the pluripotent ground state.
- Author
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Doumpas N, Söderholm S, Narula S, Moreira S, Doble BW, Cantù C, and Basler K
- Subjects
- Animals, Benzamides pharmacology, Culture Media chemistry, Culture Media pharmacology, DNA (Cytosine-5-)-Methyltransferases antagonists & inhibitors, DNA (Cytosine-5-)-Methyltransferases genetics, DNA (Cytosine-5-)-Methyltransferases metabolism, Diphenylamine analogs & derivatives, Diphenylamine pharmacology, Down-Regulation drug effects, Gene Editing, Hepatocyte Nuclear Factor 1-alpha deficiency, Hepatocyte Nuclear Factor 1-alpha metabolism, Inducible T-Cell Co-Stimulator Ligand antagonists & inhibitors, Inducible T-Cell Co-Stimulator Ligand genetics, Inducible T-Cell Co-Stimulator Ligand metabolism, Lymphoid Enhancer-Binding Factor 1 deficiency, Lymphoid Enhancer-Binding Factor 1 metabolism, Mice, Mouse Embryonic Stem Cells cytology, Mouse Embryonic Stem Cells metabolism, Octamer Transcription Factor-3 genetics, Octamer Transcription Factor-3 metabolism, Pyridines pharmacology, Pyrimidines pharmacology, RNA Interference, RNA, Small Interfering metabolism, Transcription Factor 7-Like 1 Protein deficiency, Transcription Factor 7-Like 1 Protein metabolism, Transcription Factor 7-Like 2 Protein deficiency, Transcription Factor 7-Like 2 Protein metabolism, Transcription Factors antagonists & inhibitors, Transcription Factors genetics, Transcription Factors metabolism, beta Catenin deficiency, beta Catenin genetics, AIRE Protein, Cell Self Renewal drug effects, Hepatocyte Nuclear Factor 1-alpha genetics, Lymphoid Enhancer-Binding Factor 1 genetics, Transcription Factor 7-Like 1 Protein genetics, Transcription Factor 7-Like 2 Protein genetics
- Abstract
Mouse embryonic stem cells (mESCs) can be maintained in vitro in defined N2B27 medium supplemented with two chemical inhibitors for GSK3 and MEK (2i) and the cytokine leukemia inhibitory factor (LIF), which act synergistically to promote self-renewal and pluripotency. Here, we find that genetic deletion of the four genes encoding the TCF/LEF transcription factors confers mESCs with the ability to self-renew in N2B27 medium alone. TCF/LEF quadruple knockout (qKO) mESCs display dysregulation of several genes, including Aire, Dnmt3l, and IcosL, located adjacent to each other within a topologically associated domain (TAD). Aire, Dnmt3l, and IcosL appear to be regulated by TCF/LEF in a β-catenin independent manner. Moreover, downregulation of Aire and Dnmt3l in wild-type mESCs mimics the loss of TCF/LEF and increases mESC survival in the absence of 2iL. Hence, this study identifies TCF/LEF effectors that mediate exit from the pluripotent state., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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