Liposome-mediated RNA transfection should be used with caution
Liposome-mediated RNA transfection should be used with caution
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DOI：
10.1261/rna.191706
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发表时间：
2006-10-01
期刊：
影响因子：
4.5
通讯作者：
Osborne, H. Beverley
中科院分区：
文献类型：
--
作者：
Barreau, Carine;Dutertre, Stephanie;Osborne, H. Beverley
脂质体介导的RNA转染法在研究报告基因在哺乳动物细胞中的代谢方面显示出许多优点。该方法也被广泛应用于siRNAs的转染。在这里，我们描述的结果表明，通过脂质体导入HeLa细胞的报告mRNAs没有呈现预期的行为。也就是说，报告mRNAs的稳定性与AUUA不稳定元件、Poly(A)尾巴或甚至5‘甲基化帽的存在或不存在无关。共聚焦显微镜显示，脂质体介导的荧光RNA存在于离散的颗粒中。这些观察结果表明，当使用脂质体介导RNA转染时，需要进行一些对照实验，并讨论了可能的后果。
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.