Generation and Analysis of Cartilage-Specific CCN2 Overexpression in Transgenic Mice.
Generation and Analysis of Cartilage-Specific CCN2 Overexpression in Transgenic Mice.
复制标题
转基因小鼠中软骨特异性 CCN2 过表达的生成和分析。
DOI：
10.1007/978-1-4939-6430-7_32
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
and Takigawa M
中科院分区：
文献类型：
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作者：
Hattori T;Itoh S;and Takigawa M
基因编辑技术的最新进展为提高我们对生物体分子功能的理解提供了强大的影响。在这里，我们描述了我们的方法来产生转基因过表达小鼠模型，该方法涉及使用组织特异性启动子来分析特定组织中的某些分子。本章中描述的方案使用Col 2a 1启动子-增强子，其已知用于驱动软骨中的特异性和强转基因表达，并且基于我们的几项研究，这些研究显示结缔组织生长因子（CCN 2）在软骨-骨发育和关节软骨维持中的积极作用。这些小鼠表现出强烈加速的软骨内骨化，导致骨延长增强，以及对年龄相关性关节退行性变的抵抗力。该方案还描述了如何通过使用从CCN 2过表达软骨中分离的软骨细胞来分析这些现象的分子机制。
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.