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- Title
Nanopore Sequencing Accurately Identifies the Mutagenic DNA Lesion O<sup>6</sup>‐Carboxymethyl Guanine and Reveals Its Behavior in Replication.
- Authors
Wang, Yu; Patil, Kiran M.; Yan, Shuanghong; Zhang, Panke; Guo, Weiming; Wang, Yuqin; Chen, Hong‐Yuan; Gillingham, Dennis; Huang, Shuo
- Abstract
O6‐carboxymethylguanine (O6‐CMG) is a highly mutagenic alkylation product of DNA, triggering transition mutations relevant to gastrointestinal cancer. However, precise localization of a single O6‐CMG with conventional sequencing platforms is challenging. Here nanopore sequencing (NPS), which directly senses single DNA bases according to their physiochemical properties, was employed to detect O6‐CMG. A unique O6‐CMG signal was observed during NPS and a single‐event call accuracy of >95 % was achieved. Moreover, O6‐CMG was found to be a replication obstacle for Phi29 DNA polymerase (Phi29 DNAP), suggesting this lesion could cause DNA sequencing biases in next generation sequencing (NGS) approaches.
- Subjects
GASTROINTESTINAL cancer treatment; METHYLGUANINE; CARBOXYMETHYL compounds; NUCLEOTIDE sequencing; DNA alkylation
- Publication
Angewandte Chemie, 2019, Vol 131, Issue 25, p8520
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.201902521