Nanopore Sequencing Accurately Identifies the Mutagenic DNA Lesion O6 -Carboxymethyl Guanine and Reveals Its Behavior in Replication.
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.
['Wang|Yu|Y|0000-0002-2065-8059', 'Patil|Kiran M|KM|0000-0002-9887-128X', 'Yan|Shuanghong|S|0000-0002-3292-2830', 'Zhang|Panke|P|0000-0001-8562-9972', 'Guo|Weiming|W|0000-0001-9223-0820', 'Wang|Yuqin|Y|0000-0002-9311-241X', 'Chen|Hong-Yuan|HY|', 'Gillingham|Dennis|D|0000-0002-3672-8699', 'Huang|Shuo|S|0000-0001-6133-7027']
D004247:DNA D004259:DNA-Directed DNA Polymerase D006147:Guanine D006801:Humans D015394:Molecular Structure D000081414:Nanopore Sequencing
nanopore sequencing DNA lesion gastrointestinal cancer carboxymethyl guanine