Generation of a Mouse Line Harboring a Bi-Transgene Expressing Luciferase and Tamoxifen-Activatable CreERT2 Recombinase in Cartilage
Generation of a Mouse Line Harboring a Bi-Transgene Expressing Luciferase and Tamoxifen-Activatable CreERT2 Recombinase in Cartilage
复制标题
DOI：
10.1002/dvg.22734
复制
发表时间：
2014-02-01
期刊：
影响因子：
1.5
通讯作者：
Bou-Gharios, George
中科院分区：
文献类型：
--
作者：
Lo Cascio, Leandro;Liu, Ke;Bou-Gharios, George
我们使用聚合蛋白基因增强子产生了一种转基因小鼠系（can- creer - ires - luc），表达萤火虫荧光素酶和他莫昔芬可激活的Cre重组酶（Cre- ert2）。在Acan-CreER-Ires-Luc系与Rosa26-lacZ报告基因小鼠杂交的双转基因小鼠中，检测了Cre重组酶活性的表达和效率。本系转基因的表达模式仅限于软骨从胚胎期到成体期。生长板、关节软骨、半月板纤维软骨、气管、椎间盘均可见-半乳糖苷酶染色。在先前描述的Agc1 （tm(IRES-creERT2)）小鼠系中观察到类似的染色。转基因中荧光素酶的存在使得转基因在活体动物中的表达可视化。从2周龄至8周龄的每周测量显示，在2周龄至4周龄期间膝关节发光减少，但此后稳定。在12周龄手术诱导骨关节炎后，膝关节的发光水平保持不变8周。这种Acan- creer - ires - luc小鼠系可以通过荧光素酶的表达间接监测Acan基因的转录活性，而可诱导的Cre重组酶活性有助于研究软骨中基因表达的增加或减少。创世纪1 - 10。(c) 2013 Wiley Periodicals, Inc.；
We have used an aggrecan gene enhancer to generate a transgenic murine line (Acan-CreER-Ires-Luc) expressing firefly luciferase and tamoxifen activatable Cre recombinase (Cre-ERT2). The expression and efficiency of the inducible Cre recombinase activity were tested in double transgenic mice created by crossing the Acan-CreER-Ires-Luc line with a Rosa26-lacZ reporter mouse. The expression pattern of the transgene of our line was restricted to cartilage from embryonic to adult stages. -galactosidase staining was observed in growth plate, articular cartilage, as well as fibrocartilage of meniscus, trachea, and intervertebral discs. Similar staining was observed in a previously described Agc1 (tm(IRES-creERT2)) murine line. The presence of luciferase in our transgene allows the visualization of the transgene expression in live animals. Weekly measurements from 2 to 8 weeks of age showed a reduction in luminescence in knee joints between 2 and 4 weeks of age, but stabilization thereafter. Following the surgical induction of osteoarthritis at 12 weeks of age, the level of luminescence remained the same in the knee joints for 8 weeks. This Acan-CreER-Ires-Luc murine line allows indirect monitoring of the transcriptional activity of the Acan gene via expression of luciferase, while the inducible Cre recombinase activity facilitates studies involving gain or loss of gene expression in cartilage. genesis 1-10. (c) 2013 Wiley Periodicals, Inc.