Transcriptome-wide mapping of dihydrouridine sites with D-seq

We report the development of D-Seq, a sequencing based technique for identification of dihydrouridine (D) modifications within RNA. We performed D-Seq on S. cerevisiae and find the first evidence for dihydrouridylation of endogenous mRNAs. RNA was treated with NaBH4 to modify D residues, which cause RT stops. The sites of DUS-dependent RT stops were used to identify D sites.