Hypertrophic chondrocyte-specific Col10a1 controlling elements in Cre recombinase transgenic studies

The type X collagen gene ( COL10A1 ) is specifically expressed in chondrocytes undergoing hypertrophy, which is an essential late stage of endochondral ossification during the development of long bones. We have previously localized multiple murine Col10a1 promoter-enhancer elements and used these el...

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Published in:American journal of translational research 2019-01, Vol.11 (10), p.6672-6679
Main Authors: Chen, Jinnan, Chen, Fangzhou, Bian, Huiqin, Wang, Qian, Zhang, Xiaojing, Sun, Lichun, Gu, Junxia, Lu, Yaojuan, Zheng, Qiping
Format: Article
Language:English
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Summary:The type X collagen gene ( COL10A1 ) is specifically expressed in chondrocytes undergoing hypertrophy, which is an essential late stage of endochondral ossification during the development of long bones. We have previously localized multiple murine Col10a1 promoter-enhancer elements and used these elements for transgenic studies with LacZ reporter gene or genes of interest. Here, we report two additional transgenic mouse lines in which Cre was driven by the 10 kb Col10a1 promoter/intron and the 300-bp enhancer elements respectively. Cre activity was assessed by breeding the transgenic founders onto the RosA26R genetic background and to examine its β-gal activity (blue staining) via Cre/Lox P recombination. Our results showed that, in addition to the Cre activity in hypertrophic chondrocytes, we also observed blue staining of the bone marrow and the surrounding digits when the 10 kb Col10a1 promoter/intron element was used, whereas the 300-bp enhancer element could drive Cre expression exclusively within the hypertrophic zone as demonstrated by the blue staining pattern. This is intriguing, as the 10 kb promoter covers the 300-bp enhancer element. We then further reanalyzed the LacZ transgenic mice. We did observe non-specific blue staining in 10 kb- LacZ mice but not the mice with the 300-bp enhancer. In addition, the Cre reporter construct was on a coat-color vector backbone, which enables direct visual genotyping of the transgenic mice in the FVB/N albino background. Together, our results support that the 300 bp Col10a1 enhancer provides a more efficient genetic tool to target the hypertrophic zone for studies of skeletal development and disease.
ISSN:1943-8141
1943-8141