Taniguchi Ichiro, McCloskey Asako, Ohno Mutsuhito
Institute for Virus Research, Kyoto University, Kyoto, Japan.
Methods Cell Biol. 2014;122:395-413. doi: 10.1016/B978-0-12-417160-2.00018-7.
In eukaryotes, many RNA species are transcribed, processed in the nucleus, and exported to the cytoplasm, where they are destined to function or to be further matured. Some RNAs are even reimported to the nucleus. In addition, many RNAs are localized at specific nuclear bodies before their export and/or after their nuclear reimport. To understand how RNAs are transported, Xenopus oocytes are extremely useful cells, thanks to their large size. RNA transport can be easily examined by microinjecting radioactively or fluorescently labeled RNAs into Xenopus oocytes. Mammalian cultured cells are sometimes useful by virtue of RNA-FISH technique. Here, we describe methods to analyze RNA localization and export using these cells.
在真核生物中,许多RNA种类被转录、在细胞核中加工,然后输出到细胞质,在细胞质中它们要么发挥功能,要么进一步成熟。有些RNA甚至会被重新输入细胞核。此外,许多RNA在输出之前和/或重新输入细胞核之后会定位在特定的核体中。为了了解RNA是如何运输的,非洲爪蟾卵母细胞因其体积大而成为非常有用的细胞。通过将放射性或荧光标记的RNA显微注射到非洲爪蟾卵母细胞中,可以很容易地检测RNA运输。借助RNA荧光原位杂交技术,哺乳动物培养细胞有时也很有用。在这里,我们描述了使用这些细胞分析RNA定位和输出的方法。