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目的研究CTGF-siRNA的体外转录合成及使用专门转染试剂盒将其转染至硬皮病皮肤成纤维细胞的转染效率,为将RNA干扰技术深入应用到硬皮病治疗做铺垫。方法体外培养硬皮病皮肤成纤维细胞;设计CTGF的siRNA;通过体外转录获得siRNA;将siRNA以SilencerTMsiRNALabelingKit标记;用siPORTTMAmine转染至成纤维细胞;荧光显微镜观察转染情况。结果转染率可高达85.32%。结论该方法操作简便,且可达到理想的转染率。
OBJECTIVE: To study the transfection efficiency of CTGF-siRNA in vitro and its transfection efficiency to skin fibroblasts of scleroderma using special transfection kit in order to pave the way for the further application of CTGF-siRNA in the treatment of scleroderma. METHODS: Scleroderma skin fibroblasts were cultured in vitro. SiRNA of CTGF was designed. SiRNA was obtained by in vitro transcription. SiRNA was labeled with SilencerTMsiRNALabelingKit. Transfected into fibroblasts with siPORTTMAmine. Transfection was observed by fluorescence microscopy. The results of transfection rate can be as high as 85.32%. Conclusion The method is simple and can achieve the desired transfection efficiency.