Department of Chemistry and Institute for Biophysical Dynamics, Howard Hughes Medical Institute, The University of Chicago, Chicago, IL 60637, USA; Medical Scientist Training Program, Committee on Immunology, The University of Chicago, Chicago, IL 60637, USA.
Department of Chemistry and Institute for Biophysical Dynamics, Howard Hughes Medical Institute, The University of Chicago, Chicago, IL 60637, USA.
Methods. 2019 Mar 1;156:85-90. doi: 10.1016/j.ymeth.2018.11.007. Epub 2018 Nov 22.
The post-transcriptional modification 2'-O-Methyl (2'OMe) could be present on the ribose of all four ribonucleosides, and is highly prevalent in a wide variety of RNA species, including the 5' RNA cap of viruses and higher eukaryotes, as well as internally in transfer RNA and ribosomal RNA. Recent studies have suggested that 2'OMe is also located internally in low-abundance RNA species such as viral RNA and mRNA. To profile 2'OMe on different RNA species, we have developed Nm-seq, which could identify 2'OMe sites at single base resolution. Nm-seq is particularly useful for identifying 2'OMe sites located at the 3' terminal ends of small RNAs. Here, we present an optimized protocol for Nm-seq and a protocol for applying Nm-seq to identify 2'OMe sites on small RNA 3' terminal ends.
转录后修饰 2'-O-甲基化（2'OMe）可能存在于所有四种核糖核苷的核糖上，并且在广泛的 RNA 种类中高度普遍，包括病毒和高等真核生物的 5'RNA 帽，以及转移 RNA 和核糖体 RNA 内部。最近的研究表明，2'OMe 也位于低丰度 RNA 种类（如病毒 RNA 和 mRNA）的内部。为了对不同的 RNA 种类进行 2'OMe 分析，我们开发了 Nm-seq，可以在单碱基分辨率下识别 2'OMe 位点。Nm-seq 特别适用于鉴定位于小 RNA 3'末端的 2'OMe 位点。在这里，我们提供了 Nm-seq 的优化方案以及应用 Nm-seq 鉴定小 RNA 3'末端 2'OMe 位点的方案。
