Ovchinnikov Dmitry A, van Zuylen Wendy J M, DeBats Claire E E, Alexander Kylie A, Kellie Stuart, Hume David A
Institute for Molecular Bioscience, Bldg. 80, University of Queensland, Brisbane, Queensland 4072, Australia.
J Leukoc Biol. 2008 Feb;83(2):430-3. doi: 10.1189/jlb.0807585. Epub 2007 Oct 30.
We generated double-transgenic mice carrying cointegrated tissue-specific Gal4 and Gal4 reporter transgenes to direct transgene overexpression in the mononuclear phagocyte system (MPS). A modified promoter of the Csf1r (c-fms) gene, containing a deletion of the trophoblast-specific promoter, was used to drive the expression of Gal4VP16 transcriptional activator specifically in macrophages. This module was cointegrated with a fluorescent reporter, enhanced cyan fluorescent protein (ECFP), driven by a Gal4-dependent promoter. ECFP fluorescence was first detected in forming blood islands of the yolk sac at 8 dpc, then in macrophages in the yolk sac and the embryo proper. In adult mice ECFP was detected primarily in monocytes, tissue macrophages, microglia, and dendritic cells, including Langerhans cells of the skin. Crossing of these mice to transgenics containing tagged protein under control of a Gal4-dependent promoter directed expression of that protein in mononuclear phagocytes of double-transgenic animals. The new mouse line provides a useful tool for overexpression of transgenes in cells of the myeloid lineage, while simultaneously labeling them by ECFP expression.
我们构建了携带共整合组织特异性Gal4和Gal4报告基因转基因的双转基因小鼠,以指导转基因在单核吞噬细胞系统(MPS)中过表达。使用经修饰的Csf1r(c-fms)基因启动子(缺失滋养层特异性启动子)来驱动Gal4VP16转录激活因子在巨噬细胞中特异性表达。该模块与由Gal4依赖性启动子驱动的荧光报告基因——增强型青色荧光蛋白(ECFP)共整合。在胚胎发育第8天,首先在卵黄囊形成的血岛中检测到ECFP荧光,然后在卵黄囊和胚胎本身的巨噬细胞中检测到。在成年小鼠中,ECFP主要在单核细胞、组织巨噬细胞、小胶质细胞和树突状细胞中检测到,包括皮肤的朗格汉斯细胞。将这些小鼠与在Gal4依赖性启动子控制下含有标记蛋白的转基因小鼠杂交,可使该蛋白在双转基因动物的单核吞噬细胞中定向表达。这种新的小鼠品系为在髓系细胞系中过表达转基因提供了一种有用的工具,同时通过ECFP表达对它们进行标记。