ABSTRACT

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

摘要

在过去的十年中，表观转录组学领域突出了人类 RNA 的广泛转录后修饰，包括 microRNA (miRNA)。最近的报道表明，人类 miRNA 会发生胞嘧啶甲基化。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 (hydroxy)-methyl-5-cytosine ((h)m5C) 修饰的高分辨率映射miRNA。我们的方法揭示了 miRNA 在各种序列环境中经历广泛的胞嘧啶修饰。

此外，用特异性抗体验证我们的数据揭示了人类成熟 miRNA 中的 m5C 和 hm5C 残基。BS-miRNA-seq 和 MAmBA 可能有助于 (h)m5C 在各种细胞类型和组织中的 miRNA 上的精确定位，这是理解这种修饰对 miRNA 的功能影响的关键成就。MAmBA 可在 https://github.com/flcvlr/MAmBA 下载