Dual enzyme cleavage-assisted sequencing enables genome-wide mapping of DNA damage at single-nucleotide resolution

DNA damage could compromise genome integrity and adversely affect various cellular processes, including transcription, replication, and chromatin assembly. Accurate mapping of DNA damage is crucial for elucidating their roles in both biological and pathological contexts.

In this study, we propose a dual enzyme cleavage-assisted sequencing (DEC-seq) method for genome-wide mapping of DNA damage at single-nucleotide resolution. DEC-seq integrates damage-specific glycosylase excision with restriction endonuclease cleavage to selectively enrich DNA containing damage while effectively eliminating background DNA without damage. We successfully applied DEC-seq to map uracil (U), 8-oxo-7,8-dihydroguanine (8OG), and apurinic/apyrimidinic (AP) sites in DNA at single-nucleotide resolution.