Biochemistry. 2022 Aug 16;61(16):1665-1668. doi: 10.1021/acs.biochem.2c00270. Epub 2022 Jul 25.
Herein, we detail a novel reverse-transcription (RT) assay to directly detect chemical adducts on RNA. We optimize a fluorescence quenching assay to detect RT polymerization and employ our approach to detect N-alkylation of inosine, an important post-transcriptional modification, using a phenylacrylamide as a model compound. We anticipate our approach can be expanded to identify novel reagents that form adducts with RNA and further explored to understand the relationship between RT processivity and natural post-transcriptional modifications in RNA.
在此，我们详细介绍了一种新型的逆转录（RT）测定法，可直接检测 RNA 上的化学加合物。我们优化了荧光猝灭测定法来检测 RT 聚合，并采用该方法使用苯丙烯酰胺作为模型化合物来检测肌苷的 N-烷基化，这是一种重要的转录后修饰。我们预计我们的方法可以扩展到鉴定与 RNA 形成加合物的新型试剂，并进一步探索 RT 过程性与 RNA 中天然转录后修饰之间的关系。
