Characterizing Post-transcriptional Modifications of circRNAs to Investigate Biogenesis and Translation

Recent studies have shown that circular RNAs (circRNAs) are decorated with N6-methyladenosine (m6A), a co-transcriptional modification known to participate in the regulation of many processes governing linear RNA metabolism. Nevertheless, the activity of this mark on circRNAs is still poorly understood. In order to facilitate the study of m6A-dependent regulation of these molecules, we provide protocols that enable circOme-wide detection of m6A as well as the perturbation of several components of the m6A machinery followed by assays useful to evaluate the impact of their depletion on the production and, when applicable, on the translation of circRNAs. Other modifications exist and can be explored following the same principles.