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ScxCre转基因小鼠的产生与特性分析

Generation and characterization of ScxCre transgenic mice.

作者信息

Sugimoto Yuki, Takimoto Aki, Hiraki Yuji, Shukunami Chisa

机构信息

Department of Cellular Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, 606-8507, Japan.

出版信息

Genesis. 2013 Apr;51(4):275-83. doi: 10.1002/dvg.22372. Epub 2013 Mar 13.

Abstract

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.

摘要

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。

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