Department of Transgenic Models of Diseases, Institute of Molecular Genetics of the ASCR, v.v.i., Vídeňská 1083, 14220 Prague 4, Czech Republic.
Transgenic Res. 2012 Jun;21(3):683-9. doi: 10.1007/s11248-011-9567-x. Epub 2011 Oct 22.
The epidermis is a stratified tissue composed of different keratinocyte layers that create a barrier protecting the body from external influences, pathogens, and dehydration. The barrier function is mainly achieved by its outermost layer, the stratum corneum. To create a mouse model to study pathophysiological processes in the outermost layers of the epidermis in vivo and in vitro we prepared a construct containing red fluorescent td-Tomato reporter sequence under the control of involucrin promoter and its first intron. Transgenic mice were generated by pronuclear injection and the expression and regulation of the transgene was determined by in vivo imaging and fluorescent microscopy. The promoter targeted the transgene efficiently and specifically into the outermost epidermal layers although weak expression was also found in epithelia of tongue and bladder. The regulation of expression in the epidermis, i.e. fluorescence intensity of the reporter, could be easily followed during wound healing and dermatitis. Thus, these transgenic mice carrying the tdTomato reporter could be used as a valuable tool to study impact of various genes dysregulating the epidermal barrier and to follow effects of therapeutic agents for treatment of skin diseases in vivo.
表皮是一种分层组织,由不同的角蛋白细胞层组成,形成一道屏障,保护身体免受外部影响、病原体和脱水的侵害。屏障功能主要由其最外层的角质层实现。为了建立一个研究表皮最外层生理病理过程的体内和体外小鼠模型,我们制备了一个包含受 Involucrin 启动子及其第一内含子控制的红色荧光 td-Tomato 报告序列的构建体。通过原核注射生成转基因小鼠,并通过体内成像和荧光显微镜确定转基因的表达和调控。该启动子有效地、特异性地将转基因靶向表皮的最外层,尽管在舌和膀胱的上皮中也发现了较弱的表达。在伤口愈合和皮炎过程中,很容易跟踪表皮中表达的调控,即报告基因的荧光强度。因此,这些携带 tdTomato 报告基因的转基因小鼠可作为一种有价值的工具,用于研究各种基因失调对表皮屏障的影响,并在体内跟踪治疗皮肤病的治疗剂的效果。