HELIOS NAD-Seq

This study describes HELIOS NAD-Seq, a sequencing protocol designed to identify RNAs that carry a nicotinamide adenine dinucleotide (NAD) cap at their 5' end. NAD-capped RNAs are found in many organisms and may play roles in RNA stability and host-pathogen interaction. HELIOS NAD-Seq improves on earlier methods by working with much smaller RNA inputs, showing higher specificity, and allowing many samples to be processed in parallel. Using Escherichia coli as a model, this method identified most of the previously known NAD-capped RNAs and discovered additional low-abundance transcripts. Time-course experiments across bacterial growth revealed dynamic NAD-capping patterns linked to metabolic states. These data provide a resource for studying RNA modifications and their connection to cellular physiology.