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Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
journal contribution
posted on 2020-01-09, 16:01 authored by Jiajun Wang, Meng-Yin Li, Jie Yang, Ya-Qian Wang, Xue-Yuan Wu, Jin Huang, Yi-Lun Ying, Yi-Tao LongDNA
lesions such as metholcytosine(mC), 8-OXO-guanine
(OG), 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 (K238Q), 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 K238Q 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 (mC, OG, 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.
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O GK 238Q aerolysin nanoporemodification siteslesion basesaerolysin nanoporesAerolysin Single Molecule Interface DNA lesionsnanopore electrochemistry approachlesion detectionDamaged Nucleotidesheterocomposition conditionselectrochemical readoutsDNA sequencingDNA lesion detectionK 238Qnovel K 238Qm Cmolecule interfaceDirect Quantification
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