a Goethe-University Frankfurt, Institute for Organic Chemistry and Chemical Biology , Frankfurt , Germany.
RNA Biol. 2019 Sep;16(9):1119-1132. doi: 10.1080/15476286.2019.1593093. Epub 2019 Apr 21.
Most single-molecule techniques observing RNA or require fluorescent labels that have to be connected to the RNA of interest. In recent years, a plethora of methods has been developed to achieve site-specific labelling, in many cases under near-native conditions. Here, we review chemical as well as enzymatic labelling methods that are compatible with single-molecule fluorescence spectroscopy or microscopy and show how these can be combined to offer a large variety of options to site-specifically place one or more labels in an RNA of interest. By either chemically forming a covalent bond or non-covalent hybridization, these techniques are prerequisites to perform state-of-the-art single-molecule experiments.
大多数观察 RNA 的单分子技术或需要荧光标记，这些标记必须连接到感兴趣的 RNA 上。近年来，已经开发出许多方法来实现定点标记，在许多情况下是在近天然条件下。在这里，我们回顾了与单分子荧光光谱或显微镜兼容的化学和酶标记方法，并展示了如何将这些方法结合起来，为在感兴趣的 RNA 中特异性地放置一个或多个标记提供多种选择。通过化学形成共价键或非共价杂交，这些技术是进行最先进的单分子实验的前提条件。
