Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA.
Molecules. 2021 Mar 3;26(5):1341. doi: 10.3390/molecules26051341.
The introduction of fluorophores into RNA for both in vitro and in cellulo studies of RNA function and cellular distribution is a subject of great current interest. Here I briefly review methods, some well-established and others newly developed, which have been successfully exploited to site-specifically fluorescently label interior positions of RNAs, as a guide to investigators seeking to apply this approach to their studies. Most of these methods can be applied directly to intact RNAs, including (1) the exploitation of natural posttranslational modifications, (2) the repurposing of enzymatic transferase reactions, and (3) the nucleic acid-assisted labeling of intact RNAs. In addition, several methods are described in which specifically labeled RNAs are prepared de novo.
将荧光团引入 RNA，无论是在体外还是在细胞内研究 RNA 功能和细胞分布，都是当前非常关注的课题。在这里，我简要回顾了一些已经成功应用于 RNA 内部位置特异性荧光标记的方法，这些方法有些已经建立，有些则是新开发的，以供希望将这种方法应用于自己研究的研究人员参考。这些方法中的大多数都可以直接应用于完整的 RNA，包括（1）利用天然的翻译后修饰，（2）重新利用酶转移酶反应，以及（3）利用核酸辅助完整 RNA 的标记。此外，还描述了几种从头制备特定标记 RNA 的方法。
