Sugimoto Yuki, Takimoto Aki, Hiraki Yuji, Shukunami Chisa
Department of Cellular Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 606-8507, Japan.
Genesis. 2013 Apr;51(4):275-83. doi: 10.1002/dvg.22372. Epub 2013 Mar 13.
Scleraxis (Scx) is a basic helix-loop-helix transcription factor that is a marker for the tendon/ligament cell lineage. The ∼11 kb genomic region from the mouse Scx gene locus faithfully recapitulates the endogenous Scx expression pattern in ScxGFP transgenic (Tg) mice. We have established two Tg mouse lines expressing Cre-recombinase (Cre) using this regulatory region (ScxCre-L and ScxCre-H). The specificity and efficiency of Cre recombination in these Tg lines are evaluated by crossing with Rosa-CAG-LSL-tdTomato (Ai14) or ROSA26R (R26R) reporter mice. The recombination in ScxCre-H;Ai14 mice is efficiently achieved in the endogenous Scx expression domains including the branchial arches, the syndetome, and the lateral plate mesoderm. Further analysis of ScxCre-H;Ai14;ScxGFP embryos reveal that expression of the ScxGFP transgene largely overlaps with Cre activity detected by tdTomato at embryonic day 12.5 (E12.5). In ScxCre-L;R26R or ScxCre-H;R26R neonates, Cre activity is detected in tendons, ligaments, intervertebral discs, joints, and cartilage around the chondro-tendinous/ligamentous junction, the prospective enthesis. The present results suggest that ScxCre Tg lines are useful for targeting the gene specifically in the Scx-expressing domains.
硬骨素(Scx)是一种碱性螺旋-环-螺旋转录因子,是肌腱/韧带细胞谱系的标志物。来自小鼠Scx基因座的约11 kb基因组区域忠实地再现了ScxGFP转基因(Tg)小鼠中的内源性Scx表达模式。我们利用该调控区域建立了两个表达Cre重组酶(Cre)的Tg小鼠品系(ScxCre-L和ScxCre-H)。通过与Rosa-CAG-LSL-tdTomato(Ai14)或ROSA26R(R26R)报告基因小鼠杂交,评估这些Tg品系中Cre重组的特异性和效率。在ScxCre-H;Ai14小鼠中,在内源性Scx表达域(包括鳃弓、联合中胚层和侧板中胚层)中有效地实现了重组。对ScxCre-H;Ai14;ScxGFP胚胎的进一步分析表明,在胚胎第12.5天(E12.5),ScxGFP转基因的表达与tdTomato检测到的Cre活性大部分重叠。在ScxCre-L;R26R或ScxCre-H;R26R新生小鼠中,在肌腱、韧带、椎间盘、关节以及软骨-肌腱/韧带交界处(预期的附着点)周围的软骨中检测到Cre活性。目前的结果表明,ScxCre Tg品系可用于在Scx表达域中特异性靶向基因。