Journal
CHEMICAL RESEARCH IN TOXICOLOGY
Volume 33, Issue 12, Pages 2944-2952Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.chemrestox.0c00202
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Funding
- Arkansas Biosciences Institute
- National Institutes of Environmental Health Sciences [ES029749]
- National Institute of General Medical Sciences of the National Institutes of Health [P20GM125503]
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Chemically induced DNA adducts can lead to mutations and cancer. Unfortunately, because common analytical methods (e.g., liquid chromatography-mass spectrometry) require adducts to be digested or liberated from DNA before quantification, information about their positions within the DNA sequence is lost. Advances in nanopore sequencing technologies allow individual DNA molecules to be analyzed at single-nucleobase resolution, enabling us to study the dynamic of epigenetic modifications and exposure-induced DNA adducts in their native forms on the DNA strand. We applied and evaluated the commercially available Oxford Nanopore Technology (ONT) sequencing platform for site-specific detection of DNA adducts and for distinguishing individual alkylated DNA adducts. Using ONT and the publicly available ELIGOS software, we analyzed a library of 15 plasmids containing site-specifically inserted O-6- or N-2-alkyl-2'-deoxyguanosine lesions differing in sizes and regiochemistries. Positions of DNA adducts were correctly located, and individual DNA adducts were clearly distinguished from each other.
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