Journal
ACS OMEGA
Volume 6, Issue 26, Pages 17027-17034Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsomega.1c02106
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Funding
- National Natural Science Foundation of China [91753136, 11774395, 11874414, 11674381, 11874415, 11774407, 21991133]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDB30201000, XDB28000000, XDB37010104, XDB37000000]
- Natural Science Foundation of Beijing Municipality [4181003]
- Key Research Program on Frontier Science [QYZDYSSW-SMC020, QYZDB-SSW-SLH045]
- National Key Research and Development Program [2016YFA0301500]
- National Laboratory of Biomacromolecules [2020kf02]
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This study successfully detects and characterizes single cisplatin adducts on DNA at the single-molecule level with single nucleotide spatial resolution, utilizing features such as dwell time, genome anchored current trace, and basecalling accuracy to differentiate these adducts.
Detection and characterization of an individual cisplatin adduct on a single DNA molecule is a demanding task. We explore the characteristic features of cisplatin adducts in the nanopore sequencing signal in aspects of dwell time, genome anchored current trace, and basecalling accuracy. The offset between the motor protein and the nanopore constriction region is revealed by dwell time analysis to be about 14 bases in the nanopore device as we examined. Characteristic distortions due to cisplatin adducts are illustrated in genome anchored current trace analysis, constituting the fingerprint for identification of cisplatin adduct. The sharp increase in odds ratio at the location of adducting sites provides additional feature in the detection of the adduct. By these combined methods, single cisplatin adducts can be detected with high fidelity on a single read of the DNA sequence. The study demonstrates an effective method in the detection and characterization of single cisplatin adducts on DNA at the single-molecule level and with single nucleotide spatial resolution.
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