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GEO help: Mouse over screen elements for information. |
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Status |
Public on Oct 17, 2011 |
Title |
ECAT15-2/Dppa2 transgenic rescue into ECAT15-2/Dppa2 deficient ESCs |
Organism |
Mus musculus |
Experiment type |
Expression profiling by array
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Summary |
Many transcription factors and DNA binding proteins play essential roles in the development of organs in which they are highly and/or specifically expressed. Embryonic stem cell (ESC) associated transcript (ECAT) 15-1 and ECAT15-2, also known as developmental pluripotency-associated (Dppa) 4 and Dppa2, are enriched in mouse ESCs and pre-implantation embryos, and encode homologous proteins with a common DNA binding domain known as the SAP motif. Previously, ECAT15-1 was shown to be important in the lung development, while it is dispensable in early development. In this study, we generated ECAT15-2 single and ECAT15-1/15-2 double knockout mice and found that almost all mutants, like ECAT15-1 mutants, died around birth with respiratory defects. Paradoxically, the expression of neither ECAT15-1 nor ECAT15-2 was detected in lung organogenesis. Several genes, such as Nkx2-5, Gata4, and Pitx2 were down-regulated in the ECAT15-2-null lung. On the other hand, genomic DNA of these genes showed inactive chromatin statuses in ECAT15-2-null ESCs, but not in wild-type ESCs. Chromatin immuno-precipitation (ChIP) assay revealed that ECAT15-2 binds to the regulatory region of Nkx2-5 in ESCs. These data suggest that ECAT15-2 have important roles in lung development where it is no longer expressed, by leaving epigenetic marks from earlier developmental stage.
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Overall design |
The ECAT15-2 mutant ESCs (15-1CT/CT, 15-2-/-) were established from E3.5 blasotcysts of 15-1+/CT, 15-2+/- intercross (CT allele; conditional targeted allele). Then the Flag-tagged ECAT15-2/Dppa2 transgenic vector based on piggyBac system were introduced. They were harvested on gelation coated dish with ES medium containing 15% FCS and LIF for three days. The total RNA from two wild type, two mock transduced ECAT15-2 deficient ESCs and two Flag-tagged ECAT15-2 transduced ECAT15-2 deficient ESCs were labeled with Cy3. Samples were hybridized with Whole Mouse Genome Microarray 4 x 44K (G4122F, Agilent), with the one color protocol. Data was analyzed by using GeneSpring GX11 software (Agilent).
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Contributor(s) |
Tomonori N, Megumi N |
Citation(s) |
21896782 |
Submission date |
Aug 22, 2011 |
Last update date |
Jan 12, 2017 |
Contact name |
Shinya Yamanaka |
E-mail(s) |
yamanaka@cira.kyoto-u.ac.jp
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Phone |
81-75-366-7041
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Organization name |
Center for iPS Cell Research and Applicaton (CiRA), Kyoto University
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Street address |
53 Kawahara-machi, Shogoin, Sakyo-ku
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City |
Kyoto |
ZIP/Postal code |
606-8507 |
Country |
Japan |
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Platforms (1) |
GPL7202 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Probe Name version) |
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Samples (6)
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This SubSeries is part of SuperSeries: |
GSE31584 |
Role of ECAT15-2/Dppa2 in lung development |
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Relations |
BioProject |
PRJNA155003 |
Supplementary file |
Size |
Download |
File type/resource |
GSE31582_RAW.tar |
13.2 Mb |
(http)(custom) |
TAR (of TXT) |
Processed data included within Sample table |
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