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Updated: May 6, 2026
Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
Published on: April 27, 2017
Activation of GSK3β by Sirt2 is required for early lineage commitment of mouse embryonic stem cell
Xiaoxing Si1, Wen Chen, Xudong Guo
1Clinical and Translational Research Center of Shanghai First Maternity & Infant Health Hospital, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Science and Technology, Tongji University, Shanghai, China.
Abstract:
Sirt2, a member of the NAD(+)-dependent protein deacetylase family, is increasingly recognized as a critical regulator of the cell cycle, cellular necrosis and cytoskeleton organization. However, its role in embryonic stem cells (ESCs) remains unclear. Here we demonstrate that Sirt2 is up-regulated during RA (retinoic acid)-induced and embryoid body (EB) differentiation of mouse ESCs. Using lentivirus-mediated shRNA methods, we found that knockdown of Sirt2 compromises the differentiation of mouse ESCs into ectoderm while promoting mesoderm and endoderm differentiation. Knockdown of Sirt2 expression also leads to the activation of GSK3β through decreased phosphorylation of the serine at position 9 (Ser9) but not tyrosine at position 216 (Tyr216). Moreover, the constitutive activation of GSK3β during EB differentiation mimics the effect of Sirt2 knockdown, while down-regulation of GSK3β rescues the effect of Sirt2 knockdown on differentiation. In contrast to the effect on lineage differentiation, Sirt2 knockdown and GSK3β up-regulation do not change the self-renewal state of mouse ESCs. Overall, our report reveals a new function for Sirt2 in regulating the proper lineage commitment of mouse ESCs.
Insights
Sirtuin 2 (Sirt2) regulates mouse embryonic stem cell (ESC) differentiation. Sirt2 knockdown impairs ectoderm formation while promoting mesoderm and endoderm, impacting lineage commitment via GSK3β signaling.
Area of Science:
- Stem cell biology
- Epigenetics
- Developmental biology
Background:
- Sirtuin 2 (Sirt2) is an NAD(+)-dependent deacetylase involved in cell cycle and necrosis.
- The function of Sirt2 in embryonic stem cells (ESCs) is not well understood.
- Sirt2 is implicated in regulating cellular processes, but its role in ESC differentiation requires clarification.
Purpose of the Study:
- To investigate the role of Sirt2 in mouse embryonic stem cell (ESC) differentiation.
- To elucidate the molecular mechanisms by which Sirt2 influences ESC lineage commitment.
- To determine the relationship between Sirt2 and GSK3β signaling during ESC differentiation.
Main Methods:
- Utilized lentivirus-mediated shRNA to knock down Sirt2 expression in mouse ESCs.
- Analyzed changes in cell lineage differentiation (ectoderm, mesoderm, endoderm) upon Sirt2 knockdown.
- Investigated the phosphorylation status and activation of Glycogen Synthase Kinase 3 beta (GSK3β) using Western blotting.
- Examined the effects of manipulating GSK3β activity on Sirt2 knockdown-induced differentiation changes.
Main Results:
- Sirt2 expression is upregulated during retinoic acid (RA)-induced and embryoid body (EB) differentiation of mouse ESCs.
- Knockdown of Sirt2 significantly compromises ectoderm differentiation while promoting mesoderm and endoderm differentiation.
- Sirt2 knockdown leads to Glycogen Synthase Kinase 3 beta (GSK3β) activation via decreased Ser9 phosphorylation.
- Constitutive GSK3β activation mimicked Sirt2 knockdown effects, and GSK3β inhibition rescued these effects.
- Sirt2 knockdown and GSK3β activation did not affect the self-renewal capacity of mouse ESCs.
Conclusions:
- Sirt2 plays a crucial role in regulating the lineage commitment of mouse embryonic stem cells (ESCs).
- Sirt2 influences differentiation pathways, specifically ectoderm formation, through the modulation of GSK3β activity.
- These findings reveal a novel function for Sirt2 in controlling proper ESC differentiation and lineage specification.
