Genome-Wide Single-Nucleotide Resolution Mapping of DNA Damage by Dual Enzyme Cleavage-Assisted Sequencing

Area of Science:

• Genomics
• Molecular Biology
• Biochemistry

Background:

• DNA is constantly damaged by internal and external factors, potentially disrupting essential cellular functions like replication and transcription.
• Accurate identification of DNA damage sites is vital for understanding its impact on health and disease.

Purpose of the Study:

• To introduce a novel, high-resolution method for genome-wide DNA damage mapping.
• To demonstrate the utility of this method for identifying specific DNA lesions.

Main Methods:

• Developed dual enzyme cleavage-assisted sequencing (DEC-seq), integrating glycosylase excision and restriction enzyme cleavage.
• Utilized DEC-seq to selectively enrich and map damaged DNA fragments at single-nucleotide resolution.
• Applied the method to detect uracil (U), 8-oxo-7,8-dihydroguanine (8OG), and apurinic/apyrimidinic (AP) sites.

Main Results:

• Successfully mapped U, 8OG, and AP sites genome-wide with single-nucleotide precision.
• Discovered enrichment of uracil in centromeric DNA regions.
• Demonstrated DEC-seq as an antibody-free and label-free technique.

Conclusions:

• DEC-seq provides a versatile, efficient, and high-resolution platform for mapping diverse DNA damages and modifications.
• The method overcomes limitations of damage-specific assays, offering a universal approach for genomic analysis.
• DEC-seq facilitates precise location analysis of DNA damage, aiding biological and pathological studies.