三种荧光标记的短双链RNA体外转染特性的比较

Comparison of transfection characters of three fluorescently labeled siRNA in vitro

目的:比较三种不同荧光标记的短双链RNA(siR-NA)体外转染特性.方法:利用脂质体LipofectamieTM2000转染三种荧光标记siRNA入小鼠胚胎成纤维细胞系NIH3T3,流式细胞仪观察转染效率,普通倒置荧光显微镜观察其淬灭情况,激光共聚焦显微镜观察其在细胞内的分布情况.结果:相同转染条件下三种荧光siRNA进入细胞的速度,细胞内的定位不同,转染效率不同,三种荧光标记的抗淬灭能力也不同.结论:荧光标记siRNA可以起到良好的示踪作用.用荧光标记siRNA作为无荧光siRNA的分布和转染效率参照时,要考虑到所选用荧光标记siRNA与无荧光标记siRNA的差异.

AIM： To compare the transfection character of three different fluorescently labeled small interfering RNA （siRNA） in vitro. METHODS： Three fluorescently labeled siRNA were transfected with LipofectamieTM2000 into NIH 3T3 cells for monitoring their transfection efficiency with flow cytometry, tracking their delivery with confocal microscopy and checking their photostability with fluorescent microscope. RESULTS： These three fluorescently labeled siRNA varied in intracellular distribution, transfection efficiency and quench speed. CONCLUSION： The fluorescence labeled siRNA helps to tracking the delivery of siRNA in cells. The difference between fluorescently labeled siRNA and unlabeled siRNA should be considered when the fluorescently labeled siRNA is used to study the distribution and transfection efficiency of siRNA.