文献类型：期刊文献

英文题名：Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface

作者：Wang, Jiajun[1,2];Li, Meng-Yin[1];Yang, Jie[2];Wang, Ya-Qian[2];Wu, Xue-Yuan[1];Huang, Jin[2];Ying, Yi-Lun[1];Long, Yi-Tao[1]

机构：[1]Nanjing Univ, Nanjing, Peoples R China;[2]East China Univ Sci & Technol, Shanghai, Peoples R China

年份：2020

卷号：6

期号：1

起止页码：76

外文期刊名：ACS CENTRAL SCIENCE

收录：;EI(收录号：20200508110349);WOS:【SCI-EXPANDED(收录号:WOS:000517831400011)】；

基金：This research was supported by National Natural Science Foundation of China (21834001 and 61871183, 61901171). Y.-L.Y. is sponsored by National Ten Thousand Talent Program for Young Top-Notch Talent. J.W. is sponsored by the Shanghai Sailing Program (19YF1410500) and China Postdoctoral Science Foundation (2019M651412, 2019T120309).

语种：英文

外文关键词：Gene encoding - DNA - DNA sequences - Electrochemistry - Nanopores

摘要：DNA lesions such as metholcytosine(C-m), 8-OXO-guanine ((o)G), inosine (I), etc. could cause genetic diseases. Identification of the varieties of lesion bases are usually beyond the capability of conventional DNA sequencing which is mainly designed to discriminate four bases only. Therefore, lesion detection remains a challenge due to massive varieties and less distinguishable readouts for structural variations at the molecular level. Moreover, standard amplification and labeling hardly work in DNA lesion detection. Herein, we designed a single molecule interface from the mutant aerolysin (I(238Q), whose sensing region shows high compatibility to capture and then directly convert a minor lesion into distinguishable electrochemical readouts. Compared with previous single molecule sensing interfaces, the temporal resolution of the 1(238Q aerolysin nanopore is enhanced by two orders, which has the best sensing performance in all reported aerolysin nanopores. In this work, the novel K238Q could discriminate directly at least three types of lesions (C-m, (o)G, I) without labeling and quantify modification sites under the mixed heterocomposition conditions of the oligonucleotide. Such a nanopore electrochemistry approach could be further applied to diagnose genetic diseases at high sensitivity.