Institute of Biochemistry, Department of Chemistry, University of Münster, Wilhelm-Klemm-Straße 2, D-48149 Münster, Germany.
Institute of Biochemistry, Department of Chemistry, University of Münster, Wilhelm-Klemm-Straße 2, D-48149 Münster, Germany
Open Biol. 2018 Sep;8(9). doi: 10.1098/rsob.180121.
RNA methylations play a significant regulatory role in diverse biological processes. Although the transcriptome-wide discovery of unknown RNA methylation sites is essential to elucidate their function, the development of a bigger variety of detection approaches is desirable for multiple reasons. Many established detection methods for RNA modifications heavily rely on the specificity of the respective antibodies. Thus, the development of antibody-independent transcriptome-wide methods is beneficial. Even the antibody-independent high-throughput sequencing-based methods are liable to produce false-positive or false-negative results. The development of an independent method for each modification could help validate the detected modification sites. Apart from the transcriptome-wide methods for methylation detection , methods for monitoring the presence of a single methylation at a determined site are also needed. In contrast to the transcriptome-wide detection methods, the techniques used for monitoring purposes need to be cheap, fast and easy to perform. This review considers modern approaches for site-specific detection of methylated nucleotides in RNA. We also discuss the potential of third-generation sequencing methods for direct detection of RNA methylations.
RNA 甲基化在多种生物过程中发挥着重要的调控作用。尽管全面发现未知的 RNA 甲基化位点对于阐明其功能至关重要，但出于多种原因，开发更多种类的检测方法是可取的。许多已建立的 RNA 修饰检测方法严重依赖于各自抗体的特异性。因此，开发非抗体依赖的全转录组方法是有益的。即使是非抗体依赖的基于高通量测序的方法也容易产生假阳性或假阴性结果。为每种修饰开发独立的方法有助于验证检测到的修饰位点。除了全转录组检测方法外，还需要监测特定位置单个甲基化存在的方法。与全转录组检测方法相比，用于监测目的的技术需要便宜、快速且易于操作。本文综述了用于检测 RNA 中甲基化核苷酸的位点特异性的现代方法。我们还讨论了第三代测序方法直接检测 RNA 甲基化的潜力。
