Dipartimento Di Medicina Molecolare, Sapienza Università Di Roma, Rome, Italy.
Dipartimento di Biotecnologie Cellulari Ed Ematologia, Sapienza Università di Roma, Rome, Italy.
RNA Biol. 2021 Dec;18(12):2226-2235. doi: 10.1080/15476286.2021.1927423. Epub 2021 Jun 7.
In the last decade, the field of epitranscriptomics highlighted a wide array of post-transcriptional modifications in human RNAs, including microRNAs (miRNAs). Recent reports showed that human miRNAs undergo cytosine methylation. We describe the first high-throughput NGS-based method (BS-miRNA-seq) and an analysis pipeline (MAmBA) to attain high-resolution mapping of (hydroxy)-methyl-5-cytosine ((h)m5C) modifications in human miRNAs. Our method uncovers that miRNAs undergo widespread cytosine modification in various sequence contexts.Furthermore, validation of our data with specific antibodies reveals both m5C and hm5C residues in human mature miRNAs. BS-miRNA-seq and MAmBA may contribute to the precise mapping of (h)m5C on miRNAs in various cell types and tissues, a key achievement towards the understanding of the functional implications of this modification in miRNAs. MAmBA is available for download at https://github.com/flcvlr/MAmBA.
在过去的十年中，表观转录组学领域强调了人类 RNA 中广泛存在的转录后修饰，包括 microRNAs (miRNAs)。最近的报道表明，人类 miRNAs 经历胞嘧啶甲基化。我们描述了第一个基于高通量 NGS 的方法（BS-miRNA-seq）和分析管道（MAmBA），以实现人类 miRNAs 中（羟基）-甲基-5-胞嘧啶 ((h)m5C) 修饰的高分辨率作图。我们的方法揭示了 miRNAs 在各种序列背景下经历广泛的胞嘧啶修饰。此外，用特定抗体验证我们的数据显示了人类成熟 miRNAs 中的 m5C 和 hm5C 残基。BS-miRNA-seq 和 MAmBA 可能有助于在各种细胞类型和组织中精确绘制 miRNA 上的（h)m5C，这是理解该修饰在 miRNA 中功能意义的关键成就。MAmBA 可在 https://github.com/flcvlr/MAmBA 上下载。
