 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Jul 01, 2015 |
| Title |
Dazl is required for TET-mediated reprogramming to a naïve pluripotent state (I) |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
|
| Summary |
Embryonic stem cell (ESC) cultures display a heterogeneous gene expression profile, ranging from a pristine naïve pluripotent state to a primed epiblast state. While it is known that the addition of inhibitors of GSK3β and MEK (so-called 2i conditions) push ESC cultures towards a more homogeneous naïve pluripotent state, the molecular underpinnings of this naïve transition are not completely understood. Here we demonstrate that Dazl, a RNA-binding protein previously thought to be expressed specifically in developing primordial germ cells (PGCs), marks a subpopulation of ESCs in vitro that is actively transitioning toward naïve pluripotency. In the absence of Dazl expression, ESCs fail to induce proper expression of Tet enzymes required for 5-hydroxymethylation in 2i-culture conditions. As a result, 5-hydroxymethylation of methylated cystosine residues is impaired. Indeed, we demonstrate that Tet1 and Tet2 are mRNA targets of Dazl, indicating that Dazl might play a role in protection or stabilizing these mRNA molecules. Our results provide insight in the regulation of the acquisition of naïve pluripotency and demonstrate that Dazl is required for TET-mediated cytosine hydroxymethylation in cells that are actively reprogramming to a pluripotent ground state.
|
| |
|
| Overall design |
Two independent mouse ES cell lines, Dazl-GFP and Stella-GFP, were cultured on γ-irradiated feeder MEFs in DMEM containing 15% FBS or serum-free B27N2 medium both supplemented with leukemia inhibitory factor (LIF) (Ying 2008). For the 2i experiments, 1µM MEK inhibitor PD0325901 (Axon Medchem), and 5 µM GSK3β inhibitor Kenpaullone (Tocris), were used. Cells were harvest at 0, 3 and 10 days in each condition for expression microarray analysis.
|
| |
|
| Contributor(s) |
Welling M, Chen H, Munoz J, Kester L, Junker JP, Mischerikov N, Kuijk E, Silberstein L, Kharchenko PV, Geens M, van de Velde H, van Oudenaarden A, Heck AJ, Geijsen N |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
May 19, 2015 |
| Last update date |
May 10, 2018 |
| Contact name |
Niels Geijsen |
| E-mail(s) |
n.geijsen@hubrecht.eu
|
| Organization name |
Hubrecht Institute
|
| Street address |
Uppsalalaan 8
|
| City |
Utrecht |
| ZIP/Postal code |
3584 CT |
| Country |
Netherlands |
| |
|
| Platforms (1) |
| GPL4134 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Feature Number version) |
|
| Samples (12)
|
|
| This SubSeries is part of SuperSeries: |
| GSE69357 |
Dazl is required for TET-mediated reprogramming to a naïve pluripotent state |
|
| Relations |
| BioProject |
PRJNA284413 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE69055_GeneSifterExport.txt.gz |
2.1 Mb |
(ftp)(http) |
TXT |
| GSE69055_RAW.tar |
81.9 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
| Processed data are available on Series record |
|
|
|
|
 |
Less...
More...