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Publication Record


Reconciling the different roles of Gsk3β in "naïve" and "primed" pluripotent stem cells.
Singh AM, Bechard M, Smith K, Dalton S
(2012) Cell Cycle 11: 2991-6
MeSH Terms: Animals, Cell Differentiation, Enzyme Activation, Furans, Gene Expression Regulation, Enzymologic, Glycogen Synthase Kinase 3, Glycogen Synthase Kinase 3 beta, Homeodomain Proteins, Humans, Immunoenzyme Techniques, Mice, Nanog Homeobox Protein, Phosphatidylinositol 3-Kinases, Phosphorylation, Pluripotent Stem Cells, Proto-Oncogene Proteins c-myc, Pyridines, Pyrimidines, Species Specificity, Wnt Signaling Pathway
Show Abstract · Added August 25, 2016
Signaling pathways orchestrated by PI3K/Akt, Raf/Mek/Erk and Wnt/β-catenin are known to play key roles in the self-renewal and differentiation of pluripotent stem cells. The serine/threonine protein kinase Gsk3β has roles in all three pathways, making its exact function difficult to decipher. Consequently, conflicting reports have implicated Gsk3β in promoting self-renewal, while others suggest that it performs roles in the activation of differentiation pathways. Different thresholds of Gsk3β activity also have different biological effects on pluripotent cells, making this situation even more complex. Here, we describe a further level of complexity that is most apparent when comparing "naïve" murine and "primed" human pluripotent stem cells. In naïve cells, Gsk3β activity is restrained by PI3K/Akt, but when released from inhibitory signals it antagonizes self-renewal pathways by targeting pluripotency factors such as Myc and Nanog. This situation also applies in primed cells, but, in addition, a separate pool of Gsk3β is required to suppress canonical Wnt signaling. These observations suggest that different Gsk3β-protein complexes shift the balance between naïve and primed pluripotent cells and identify fundamental differences in their cell signaling. Altogether, these findings have important implications for the mechanisms underpinning the establishment of different pluripotent cell states and for the control of self-renewal and differentiation.
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20 MeSH Terms
NANOG reporter cell lines generated by gene targeting in human embryonic stem cells.
Fischer Y, Ganic E, Ameri J, Xian X, Johannesson M, Semb H
(2010) PLoS One 5:
MeSH Terms: Cell Line, Embryonic Stem Cells, Gene Expression Regulation, Gene Targeting, Genes, Reporter, Homeodomain Proteins, Humans, Nanog Homeobox Protein
Show Abstract · Added September 19, 2017
BACKGROUND - Pluripotency and self-renewal of human embryonic stem cells (hESCs) is mediated by a complex interplay between extra- and intracellular signaling pathways, which regulate the expression of pluripotency-specific transcription factors. The homeodomain transcription factor NANOG plays a central role in maintaining hESC pluripotency, but the precise role and regulation of NANOG are not well defined.
METHODOLOGY/PRINCIPAL FINDINGS - To facilitate the study of NANOG expression and regulation in viable hESC cultures, we generated fluorescent NANOG reporter cell lines by gene targeting in hESCs. In these reporter lines, the fluorescent reporter gene was co-expressed with endogenous NANOG and responded to experimental induction or repression of the NANOG promoter with appropriate changes in expression levels. Furthermore, NANOG reporter lines facilitated the separation of hESC populations based on NANOG expression levels and their subsequent characterization. Gene expression arrays on isolated hESC subpopulations revealed genes with differential expression in NANOG(high) and NANOG(low) hESCs, providing candidates for NANOG downstream targets hESCs.
CONCLUSION/SIGNIFICANCE - The newly derived NANOG reporter hESC lines present novel tools to visualize NANOG expression in viable hESCs. In future applications, these reporter lines can be used to elucidate the function and regulation of NANOG in pluripotent hESCs.
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8 MeSH Terms