Barreau Carine, Dutertre Stéphanie, Paillard Luc, Osborne H Beverley
RNA. 2006 Oct;12(10):1790-3. doi: 10.1261/rna.191706. Epub 2006 Aug 18.
Liposome-mediated RNA transfection appears to present a number of advantages for studying the metabolism of reporter mRNAs in mammalian cells. This method is also widely used to transfect siRNAs. Here we describe results indicating that reporter mRNAs introduced into HeLa cells by liposomes do not present the expected behaviors. Namely, the stability of reporter mRNAs was independent of the presence or absence of an AUUUA instability element, a poly(A) tail, or even a 5' methylated cap. Confocal microscopy showed that fluorescent RNAs introduced by liposome-mediated transfection were present in discrete particles. These observations imply that a number of control experiments are required when using liposome to mediated RNA transfection, and the possible consequences are discussed.
脂质体介导的RNA转染对于研究哺乳动物细胞中报告基因mRNA的代谢似乎具有许多优势。该方法也广泛用于转染小干扰RNA（siRNA）。在此，我们描述的结果表明，通过脂质体导入HeLa细胞的报告基因mRNA并未表现出预期的行为。具体而言，报告基因mRNA的稳定性与是否存在AUUUA不稳定元件、聚腺苷酸（poly(A)）尾，甚至5'甲基化帽无关。共聚焦显微镜显示，脂质体介导的转染所导入的荧光RNA存在于离散颗粒中。这些观察结果表明，在使用脂质体介导RNA转染时需要进行大量对照实验，并对可能的结果进行了讨论。
