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Status |
Public on Dec 22, 2021 |
Title |
The effect of S309W mutation on SP7 genomic binding in chondrocytes |
Organism |
Mus musculus |
Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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Summary |
SP7/Osterix is a transcription factor critical for osteoblast maturation and bone formation. We identidied a missense variant (c.926C>G:p.S309W) in SP7 in a patient with a unique high turnover bone disease. Mice with the corresponding variant similarly showed a complex skeletal phenotype distinct from that of Sp7-null mice. We therefore performed ChIP-Seq in primary chondrocytes to study how the mutation alters the genomic binding of SP7.
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Overall design |
Primary mouse chondrocytes isolated from proximal tibia and distal femur of 7 day old male mice cardiomyocytes were cultured in DMEM/F12 media supplemented with 10% FBS, P/S, and 50 μg/mL ascorbic acid. Retrovirus expressing either wild-type SP7 or S309W variant of SP7 were produced with HEK-derived ECO cells and used for infecting primary chondrocytes. About 5 days post-infection, chondrocytes were harvested and cross-linked with 1% formaldehyde for genomic DNA isolation.
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Contributor(s) |
Lui JC, Baron J, Hojo H |
Citation(s) |
35121733 |
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Submission date |
May 26, 2020 |
Last update date |
Feb 23, 2022 |
Contact name |
Hironori Hojo |
E-mail(s) |
hojo@tetrapod.t.u-tokyo.ac.jp
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Phone |
+81-3-5841-1427
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Organization name |
The University of Tokyo
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Department |
Center for Disease Biology and Integrative Medicine
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Lab |
Clinical Biotechnology
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Street address |
7-3-1 Hongo
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City |
Bunkyo-ku |
State/province |
Tokyo |
ZIP/Postal code |
113-8655 |
Country |
Japan |
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Platforms (1) |
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Samples (3) |
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Relations |
BioProject |
PRJNA635142 |
SRA |
SRP264822 |