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Generation of a mouse line harboring a bi‐transgene expressing luciferase and tamoxifen‐activatable creER T2 recombinase in cartilage
Author(s) -
Cascio Leandro Lo,
Liu Ke,
Nakamura Hiro,
Chu Grace,
Lim Ngee Han,
Chanalaris Anastasios,
Saklatvala Jeremy,
Nagase Hideaki,
BouGharios George
Publication year - 2014
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/dvg.22734
Subject(s) - luciferase , cre recombinase , transgene , microbiology and biotechnology , biology , genetically modified mouse , cartilage , reporter gene , aggrecan , transgenesis , cell culture , gene expression , transfection , gene , pathology , anatomy , medicine , osteoarthritis , embryogenesis , articular cartilage , biochemistry , embryo , genetics , alternative medicine , reproductive biology
Summary 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‐ER T2 ). 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. β‐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 52:110–119. © 2013 Wiley Periodicals, Inc.