Hartwich Heiner, Satheesh Somisetty V, Nothwang Hans Gerd
Department of Neurogenetics, Institute of Biology and Environmental Sciences, Carl von Ossietzky University, Oldenburg, Germany.
BMC Res Notes. 2012 Jun 14;5:296. doi: 10.1186/1756-0500-5-296.
Targeted genetic modification in the mouse becomes increasingly important in biomedical and basic science. This goal is most often achieved by use of the Cre/loxP system and numerous Cre-driver mouse lines are currently generated. Their initial characterization requires reporter mouse lines to study the in vivo spatiotemporal activity of Cre.
Here, we report a dual fluorescence reporter mouse line, which switches expression from the red fluorescent protein mCherry to eGFP after Cre-mediated recombination. Both fluorescent proteins are expressed from the ubiquitously active and strong CAGGS promoter. Among the founders, we noticed a pink mouse line, expressing high levels of the red fluorescent protein mCherry throughout the entire body. Presence of mCherry in the living animal as well as in almost all organs was clearly visible without optical equipment. Upon Cre-activity, mCherry expression was switched to eGFP, demonstrating functionality of this reporter mouse line.
The pink mouse presented here is an attractive novel reporter line for fluorescence-based monitoring of Cre-activity. The high expression of mCherry, which is visible to the naked eye, facilitates breeding and crossing, as no genotyping is required to identify mice carrying the reporter allele. The presence of two fluorescent proteins allows in vivo monitoring of recombined and non-recombined cells. Finally, the pink mouse is an eye-catching animal model to demonstrate the power of transgenic techniques in teaching courses.
在生物医学和基础科学领域,小鼠的靶向基因修饰变得越来越重要。这一目标通常通过使用Cre/loxP系统来实现,目前已经产生了许多Cre驱动的小鼠品系。对它们的初步表征需要报告基因小鼠品系来研究Cre在体内的时空活性。
在此,我们报告了一种双荧光报告基因小鼠品系,在Cre介导的重组后,其表达从红色荧光蛋白mCherry切换为eGFP。两种荧光蛋白均由广泛活跃且强大的CAGGS启动子表达。在这些奠基小鼠中,我们注意到一个粉色小鼠品系,其在全身表达高水平的红色荧光蛋白mCherry。在没有光学设备的情况下,活体动物以及几乎所有器官中mCherry的存在都清晰可见。在Cre激活后,mCherry的表达切换为eGFP,证明了该报告基因小鼠品系的功能。
这里展示的粉色小鼠是一种用于基于荧光监测Cre活性的极具吸引力的新型报告基因品系。肉眼可见的mCherry的高表达便于繁殖和杂交,因为无需进行基因分型来鉴定携带报告基因等位基因的小鼠。两种荧光蛋白的存在允许在体内监测重组和未重组的细胞。最后,粉色小鼠是一种引人注目的动物模型,可用于在教学课程中展示转基因技术的强大之处。