Department of Chemistry and Centre for Research on Biomolecular Interactions, York University, 4700 Keele Street, Toronto, ON M3J 1P3, Canada.
J Am Chem Soc. 2022 Apr 6;144(13):5723-5727. doi: 10.1021/jacs.2c00670. Epub 2022 Mar 22.
Chemical modifications regulate the fate and function of cellular RNAs. Newly developed sequencing methods have allowed a deeper understanding of the biological role of RNA modifications; however, the vast majority of post-transcriptional modifications lack a well-defined sequencing method. Here, we report a photo-oxidative sequencing (PhOxi-seq) approach for guanosine -methylation, a common methylation mark seen in -methylguanosine (mG) and ,-dimethylguanosine (mG). Using visible light-mediated organic photoredox catalysis, mG and mG are chemoselectively oxidized in the presence of canonical RNA nucleosides, which results in a strong mutation signature observed during sequencing. PhOxi-seq was demonstrated on various tRNAs and rRNA to reveal -methylation with excellent response and markedly improved read-through at mG sites.
化学修饰调节细胞 RNA 的命运和功能。新开发的测序方法使人们能够更深入地了解 RNA 修饰的生物学作用；然而，绝大多数转录后修饰缺乏明确的测序方法。在这里，我们报告了一种用于鸟苷 -甲基化的光氧化测序（PhOxi-seq）方法，鸟苷 -甲基化是 -甲基鸟苷（mG）和 -二甲基鸟苷（mG）中常见的甲基化标记。使用可见光介导的有机光氧化还原催化，在标准 RNA 核苷存在的情况下，mG 和 mG 被选择性地化学氧化，这导致在测序过程中观察到强烈的突变特征。PhOxi-seq 在各种 tRNA 和 rRNA 上的演示表明，在 mG 位点具有优异的响应性和明显改善的通读性的 -甲基化。
