Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, PO Box 210172, Cincinnati, OH 45221-0172, United States.
Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, PO Box 210172, Cincinnati, OH 45221-0172, United States.
Methods. 2019 Mar 1;156:128-138. doi: 10.1016/j.ymeth.2018.10.012. Epub 2018 Oct 24.
Research into post-transcriptional processing and modification of RNA continues to speed forward, as their ever-emerging role in the regulation of gene expression in biological systems continues to unravel. Liquid chromatography tandem mass spectrometry (LC-MS/MS) has proven for over two decades to be a powerful ally in the elucidation of RNA modification identity and location, but the technique has not proceeded without its own unique technical challenges. The throughput of LC-MS/MS modification mapping experiments continues to be impeded by tedious and time-consuming spectral interpretation, particularly during for the analysis of complex RNA samples. RNAModMapper was recently developed as a tool to improve the interpretation and annotation of LC-MS/MS data sets from samples containing post-transcriptionally modified RNAs. Here, we delve deeper into the methodology and practice of RNAModMapper to provide greater insight into its utility, and remaining hurdles, in current RNA modification mapping experiments.
RNA 的转录后加工和修饰研究继续加速，因为它们在生物系统中基因表达调控中的作用不断被揭示。液相色谱串联质谱（LC-MS/MS）在确定 RNA 修饰的身份和位置方面已经证明是一种强大的工具，但其发展并非没有自身独特的技术挑战。LC-MS/MS 修饰图谱实验的通量仍然受到繁琐且耗时的光谱解释的阻碍，尤其是在分析复杂的 RNA 样品时。最近开发了 RNAModMapper 作为一种工具，以改善包含转录后修饰 RNA 的样品的 LC-MS/MS 数据集的解释和注释。在这里，我们深入研究了 RNAModMapper 的方法和实践，以更深入地了解其在当前 RNA 修饰图谱实验中的效用和仍然存在的障碍。
