Chapter title |
Generation and Analysis of Cartilage-Specific CCN2 Overexpression in Transgenic Mice.
|
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Chapter number | 32 |
Book title |
CCN Proteins
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Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6430-7_32 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6428-4, 978-1-4939-6430-7
|
Authors |
Takako Hattori Ph.D., Shinsuke Itoh, Masaharu Takigawa D.D.S., Ph.D., Takako Hattori, Masaharu Takigawa |
Editors |
Masaharu Takigawa |
Abstract |
Recent progress in gene-editing technology has provided a strong impact for improved our understanding of molecular functions in living organisms. Here we describe our method to generate transgene-overexpressing mouse models, which method involves the use of tissue-specific promoters for analyzing a certain molecule (s) in special tissues. The protocol described in this chapter uses the Col2a1 promoter-enhancer, which is known for driving specific and strong transgene expression in cartilage and is based on several of our studies showing a positive role of the connective tissue growth factor (CCN2) in cartilage-bone development and maintenance of articular cartilage. These mice show strongly accelerated endochondral ossification resulting in enhanced bone elongation, as well as resistance to age-related articular degeneration. This protocol also describes how to analyze the molecular mechanisms of these phenomena by use of chondrocytes isolated from CCN2-overexpressing cartilage. |
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