CompasSeq: epitranscriptome-wide percentage assessment of metabolite-capped RNA at the transcript resolution

Recent studies have revealed that nucleotide-containing metabolites, including nicotinamide adenine dinucleotide (NAD), flavin adenine dinucleotide (FAD), 3'-dephospho-coenzyme A (dpCoA), uridine diphosphate glucose (UDP-Glc), and uridine diphosphate N-acetyl glucosamine (UDP-GlcNAc), can be incorporated into the RNA 5'-terminus as noncanonical initiating nucleotides (NCIN), resulting in NCIN-capped RNA (NCIN-RNA). Overall design: We subjected 20 µg total RNA from mouse liver mixed with 0.014 ng spike-in RNAs with ascending ratios of NAD-capped forms 0% (i.e., 100% m7G-capped), 1%, 5%, 10%, and 20% to CompasSeq, followed by CompassAnalyzer analysis of NCIN-RNA profiles.