Mori Mikiro, Gargowitsch Laetitia, Bornert Jean-Marc, Garnier Jean-Marie, Mark Manuel, Chambon Pierre, Metzger Daniel
Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS UMR7104/INSERM U964, Collège de France, Université de Strasbourg, Illkirch Cedex, France.
Genesis. 2012 Nov;50(11):828-32. doi: 10.1002/dvg.22044. Epub 2012 Jul 11.
To generate temporally controlled site-specific somatic mutations in the mouse eye pigment epithelium, we generated a TRP1-Cre-ER(T2) transgenic mouse line that expresses the tamoxifen-dependent Cre-ER(T2) recombinase under the control of the tyrosinase-related protein 1 (TRP1) promoter. Cre-ER(T2) transcripts were readily detected in the retinal pigment epithelium (RPE), and tamoxifen treatment of adult TRP1-Cre-ER(T2) transgenic mice induced efficient excision of floxed DNA in patches of RPE cells, in numerous epithelial cells of the iris and ciliary body, and in very few cells of the neural retina. Importantly, no excision was detected in any cells in the absence of tamoxifen treatment. Thus, the TRP1-Cre-ER(T2) mouse line provides a powerful tool to study in vivo gene functions in the mouse eye pigment epithelium.
为了在小鼠眼色素上皮细胞中产生时间可控的位点特异性体细胞突变,我们构建了一个TRP1-Cre-ER(T2)转基因小鼠品系,该品系在酪氨酸酶相关蛋白1(TRP1)启动子的控制下表达他莫昔芬依赖性Cre-ER(T2)重组酶。在视网膜色素上皮(RPE)中很容易检测到Cre-ER(T2)转录本,用他莫昔芬处理成年TRP1-Cre-ER(T2)转基因小鼠可诱导RPE细胞斑块、虹膜和睫状体的许多上皮细胞以及神经视网膜的极少数细胞中loxP侧翼DNA的有效切除。重要的是,在没有他莫昔芬处理的情况下,未在任何细胞中检测到切除。因此,TRP1-Cre-ER(T2)小鼠品系为研究小鼠眼色素上皮细胞中的体内基因功能提供了一个强大的工具。