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| Status |
Public on Feb 27, 2012 |
| Title |
Transcriptional Activation by Oct4 Is Sufficient for the Maintenance and Induction of Pluripotency |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Oct4 is an essential regulator of embryonic stem (ES) cell pluripotency in vivo and in vitro, as well as a key mediator of the reprogramming of somatic cells to form induced pluriopotent stem (iPS) cells. It is not known whether activation and/or repression of specific genes by Oct4 is relevant to these functions. Here we show that fusion proteins containing the coding sequence of Oct4 or Xlpou91 (the Xenopus homologue of Oct4) fused to activating regions, but not those fused to repressing regions, behave as Oct4, suppressing differentiation and promoting maintenance of undifferentiated phenotypes in vivo and in vitro. An Oct4 activation domain fusion supported ES cell self-renewal in vitro at lower concentrations than required for Oct4 while alleviating the ordinary requirement for the cytokine LIF. At still lower levels of the fusion, LIF-dependence was restored. We conclude that the necessary and sufficient function of Oct4 in promoting pluripotency is to activate specific target genes.
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| Overall design |
Two independent clones from each cell line were used as biological replications. Cy3-CTP-labelled sample targets were prepared from 2.5 μg of total RNA using the Low RNA Input Fluorescent Linear Amplification Kit (Agilent). Cy5-CTP-labelled reference target was produced from 2.5 μg of Stratagene Universal Mouse Reference RNA. Purified target RNA were hybridized to the NIA Mouse 44K Microarray v3.0 (whole genome 60-mer oligo arrays, Agilent Technology, design ID 015087) (Carter et al., 2005) according to the manufacturer's protocol (Two-Colour Microarray-Based Gene Expression Analysis Protocol, Version 5.0.1). Slides were scanned with an Agilent DNA Microarray Scanner (model G2505-64120) at 100% PMT for the Cy3 channel and 10% PMT for the Cy5 channel with a scan resolution of 5µm. Data was analyzed using NIA Array Analysis (Sharov et al., 2005) using standard statistical conditions (FDR<0.05, 2-fold expression levels change) to unveil genes with changes in expression levels between the samples and controls.
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| Contributor(s) |
Hammachi F, Morrison G, Livigni A, Sharov A, O’Malley J, Kaji K, Narayan S, Papapetrou E, Ptashne M, Ko MS, Brickman JM |
| Citation(s) |
21083502, 22832160 |
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| Submission date |
Oct 06, 2010 |
| Last update date |
Aug 20, 2015 |
| Contact name |
Minoru S.H. Ko |
| E-mail(s) |
kom@mail.nih.gov
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| Phone |
410-558-8359
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| Organization name |
NIH
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| Department |
National Institute on Aging
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| Lab |
Lab of Genetics
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| Street address |
251 Bayview Blvd, Suite 100, 10C
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| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21224 |
| Country |
USA |
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| Platforms (1) |
| GPL6867 |
NIA Mouse 44K Microarray v3.0 (Whole Genome 60-mer Oligo) [Agilent 015087] |
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| Samples (18)
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| GSM605610 |
Oct4 VP2 Lif- 48h, rep2 |
| GSM605611 |
Oct4 Lif- 48h, rep1 |
| GSM605612 |
Oct4 Lif- 48h, rep2 |
| GSM605613 |
Oct4 VP2 Lif- 120h, rep1 |
| GSM605614 |
Oct4 VP2 Lif- 120h, rep2 |
| GSM605615 |
Oct4 Lif- 120h, rep1 |
| GSM605616 |
Oct4 Lif- 120h, rep2 |
| GSM605617 |
ZHBTc4 Lif- 48h, rep1 |
| GSM605618 |
ZHBTc4 Lif- 120h, rep1 |
| GSM605619 |
ZHBTc4 Lif+ 24h, rep1 |
| GSM605620 |
ZHBTc4 Lif+ 24h, rep2 |
| GSM605621 |
ZHBTc4 Lif- 48h, rep2 |
| GSM605622 |
ZHBTc4 Lif- 120h, rep2 |
| GSM605623 |
Oct4 Lif+ 24h, rep1 |
| GSM605624 |
Oct4 Lif+ 24h, rep2 |
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| Relations |
| BioProject |
PRJNA132631 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE24563_RAW.tar |
204.7 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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