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| Status |
Public on Apr 28, 2016 |
| Title |
Sp7/Osterix is restricted to bone-forming vertebrates where it acts as a Dlx co-factor in osteoblast specification [ChIP-Seq 2] |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
To identify Sp7’s molecular interactions in vivo, we used gene-targeting strategies to generate an Sp7-Biotin-3xFLAG knock-in mouse (Sp7-BioFL mouse). This approach appends a biotin (Bio) recognition motif and three copies of the FLAG (FL) epitope at the C-terminus of the Sp7 protein. To validate this knockin system, we compared ChIP-seq results in the MC3T3E1 osteoblast cell line, introducing Sp7 forms that were epitope-tagged at either the N- (N-terminal FLAG tag; MC3_FLAG-Sp7) or C-terminus (C-terminal Biotin-FLAG tag, as in the in vivo targeted allele; MC3_Sp7-BioFLAG). These data show an extensive overlap, suggesting that FLAG tagging at different positions gives comparable outcomes. Second, we examined ChIP on wild-type calvarial primary osteoblasts (POB) using an anti-Sp7 antibody (POB_Sp7Ab-ChIP). The majority of Sp7-Ab ChIP-seq peaks (95%) overlapped with the larger set identified by Sp7-BioFL ChIP-seq.
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| Overall design |
ChIP-seq experiments with primary osteoblasts and a osteoblast cell line were performed
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| |
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| Contributor(s) |
Hojo H, Ohba S, He X, Lai LP, McMahon AP |
| Citation(s) |
27134141 |
| Submission date |
Apr 13, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Hironori Hojo |
| E-mail(s) |
hojo@tetrapod.t.u-tokyo.ac.jp
|
| Phone |
+81-3-5841-1427
|
| Organization name |
The University of Tokyo
|
| Department |
Center for Disease Biology and Integrative Medicine
|
| Lab |
Clinical Biotechnology
|
| Street address |
7-3-1 Hongo
|
| City |
Bunkyo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
113-8655 |
| Country |
Japan |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
|
| Samples (5)
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| This SubSeries is part of SuperSeries: |
| GSE76187 |
Sp7/Osterix is restricted to bone-forming vertebrates where it acts as a Dlx co-factor in osteoblast specification |
|
| Relations |
| BioProject |
PRJNA318364 |
| SRA |
SRP073243 |
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