siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model
siRNA silencing of keratinocyte-specific GFP expression in a transgenic mouse skin model
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DOI：
10.1038/gt.2009.62
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发表时间：
2009-08-01
期刊：
影响因子：
5.1
通讯作者：
Kaspar, R. L.
中科院分区：
文献类型：
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作者：
Gonzalez-Gonzalez, E.;Ra, H.;Kaspar, R. L.
小干扰 RNA (siRNA) 可以被设计为特异性、有效地靶向和沉默突变等位基因，对相应的野生型等位基因表达影响很小或没有影响，从而为治疗干预提供了机会。尽管一些 siRNA 已进入临床试验，但 siRNA 疗法作为新药物类别的开发将需要开发改进的递送技术。在本研究中，开发了报告小鼠模型（转基因点击甲虫荧光素酶/人源化怪物绿色荧光蛋白），以实现 siRNA 递送至皮肤的研究；在这种转基因小鼠中，绿色荧光蛋白报告基因的表达仅限于表皮。通过组织学、实时定量聚合酶链反应和使用双轴共聚焦荧光显微镜的活体成像测量，皮内注射靶向报告基因的 siRNA 导致局部治疗区域的绿色荧光蛋白表达显着减少。这些结果表明，这种转基因小鼠皮肤模型与体内成像相结合，将有助于开发高效且“患者友好”的 siRNA 递送技术，并应有助于将基于 siRNA 的疗法转化为临床治疗皮肤疾病。基因治疗 (2009) 16, 963-972; doi：10.1038/gt.2009.62； 2009 年 5 月 28 日在线发布
Small interfering RNAs (siRNAs) can be designed to specifically and potently target and silence a mutant allele, with little or no effect on the corresponding wild-type allele expression, presenting an opportunity for therapeutic intervention. Although several siRNAs have entered clinical trials, the development of siRNA therapeutics as a new drug class will require the development of improved delivery technologies. In this study, a reporter mouse model (transgenic click beetle luciferase/humanized monster green fluorescent protein) was developed to enable the study of siRNA delivery to skin; in this transgenic mouse, green fluorescent protein reporter gene expression is confined to the epidermis. Intradermal injection of siRNAs targeting the reporter gene resulted in marked reduction of green fluorescent protein expression in the localized treatment areas as measured by histology, real-time quantitative polymerase chain reaction and intravital imaging using a dual-axes confocal fluorescence microscope. These results indicate that this transgenic mouse skin model, coupled with in vivo imaging, will be useful for development of efficient and 'patient-friendly' siRNA delivery techniques and should facilitate the translation of siRNA-based therapeutics to the clinic for treatment of skin disorders. Gene Therapy (2009) 16, 963-972; doi: 10.1038/gt.2009.62; published online 28 May 2009