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首页> 外文期刊>Molecular and Cellular Biology >Exploiting the Keratin 17 Gene Promoter To Visualize Live Cells in Epithelial Appendages of Mice
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Exploiting the Keratin 17 Gene Promoter To Visualize Live Cells in Epithelial Appendages of Mice

机译：利用角蛋白17基因启动子可视化小鼠上皮附属物中的活细胞

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摘要

Keratin genes afford, given their large number (>50) and differential regulation, a unique opportunity to study the mechanisms underlying specification and differentiation in epithelia of higher metazoans. Moreover, the small size and regulation in cis of many keratin genes enable the use of their regulatory sequence to achieve targeted gene expression in mice. Here we show that 2 kilobases of 5′ upstream region from the mouse keratin 17 gene (mK17) confers expression of green fluorescent protein (GFP) in major epithelial appendages of transgenic mice. Like that of mK17, onset of [mK17 5′]-GFP reporter expression coincides with the appearance of ectoderm-derived epithelial appendages during embryonic development. In adult mice, [mK17 5′]-GFP is appropriately regulated within hair, nail, glands, and oral papilla. Tracking of GFP fluorescence allows for the visualization of growth cycle-related changes in hair follicles, and the defects engendered by the hairless mutation, in live skin tissue. Deletion of an internal 48-bp interval, which encompasses a Gli-responsive element, from this promoter results in loss of GFP fluorescence in most appendages in vivo, suggesting that sonic hedgehog participates in K17 regulation. The compact mK17 gene promoter provides a novel tool for appendage-preferred gene expression and manipulation in transgenic mice.
机译：鉴于角蛋白基因的数量众多（> 50）和差异调节，它们提供了独特的机会来研究高等子生上皮细胞的规格和分化机制。此外，许多角蛋白基因的小尺寸和 cis 的调控使其能够利用其调控序列在小鼠中实现靶向的基因表达。在这里，我们显示了来自小鼠角蛋白17基因（ mK17 ）5'上游区域的2 kb赋予转基因小鼠主要上皮附件中绿色荧光蛋白（GFP）的表达。像 mK17 一样，[ mK17 5']-GFP报告基因的表达与胚胎发育过程中外胚层上皮附件的出现相吻合。在成年小鼠中，[ mK17 5']-GFP在头发，指甲，腺体和口腔乳头中得到适当调节。跟踪GFP荧光可以观察到活的皮肤组织中与毛囊生长周期相关的变化以及 lessless 突变引起的缺陷。从该启动子中删除一个包含Gli应答元件的内部48 bp间隔会导致体内大多数附件中的GFP荧光丧失，这表明声波刺猬参与了K17调控。紧凑型 mK17 基因启动子为在转基因小鼠中以附肢优选的基因表达和操纵提供了一种新颖的工具。

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