Wang Yu, Patil Kiran M, Yan Shuanghong, Zhang Panke, Guo Weiming, Wang Yuqin, Chen Hong-Yuan, Gillingham Dennis, Huang Shuo
School of Chemistry and Chemical Engineering, State Key Laboratory of Analytical Chemistry for Life Science and Collaborative Innovation Center of Chemistry for Life Sciences, Nanjing University, 210023, China.
Department of Chemistry, University of Basel, CH-4056, Basel, Switzerland.
Angew Chem Int Ed Engl. 2019 Jun 17;58(25):8432-8436. doi: 10.1002/anie.201902521. Epub 2019 May 13.
O -carboxymethylguanine (O -CMG) is a highly mutagenic alkylation product of DNA, triggering transition mutations relevant to gastrointestinal cancer. However, precise localization of a single O -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 O -CMG. A unique O -CMG signal was observed during NPS and a single-event call accuracy of >95 % was achieved. Moreover, O -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.
O-羧甲基鸟嘌呤（O-CMG）是DNA的一种高致突变性烷基化产物，可引发与胃肠道癌相关的转换突变。然而，使用传统测序平台精确定位单个O-CMG具有挑战性。在此，采用根据单个DNA碱基的物理化学性质直接进行检测的纳米孔测序（NPS）来检测O-CMG。在纳米孔测序过程中观察到了独特的O-CMG信号，单事件调用准确率达到了95%以上。此外，发现O-CMG是Phi29 DNA聚合酶（Phi29 DNAP）的复制障碍，这表明该损伤可能会在下一代测序（NGS）方法中导致DNA测序偏差。
