Novel chemical approach for comprehensive pseudouridine sequencing across diverse RNA types and mammalian species

This knowledge gap partially stems from the absence of accurate and unbiased detection methods. We systematically screened chemical reagents for pseudouridine-specific alkylation and cyclization that lead to mutations upon reverse transcription.

This mutation-based method, PUM-seq, is capable of mapping Ψ sites as mutation signatures across the transcriptome at single-nucleotide resolution. PUM-seq efficiently detected all known Ψ sites in human rRNA, tRNA, mRNA and all other RNA species with high sensitivity.