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Identifying DNA mismatches at single-nucleotide resolution by probing individual surface potentials of DNA-capped nanoparticles

Authors
 Hyungbeen Lee  ;  Sang Won Lee  ;  Gyudo Lee  ;  Wonseok Lee  ;  Kihwan Nam  ;  Jeong Hoon Lee  ;  Kyo Seon Hwang  ;  Jaemoon Yang  ;  Hyeyoung Lee  ;  Sangsig Kim  ;  Sang Woo Lee  ;  Dae Sung Yoon 
Citation
 Nanoscale, Vol.10(2) : 538-547, 2018 
Journal Title
 Nanoscale 
ISSN
 2040-3364 
Issue Date
2018
Abstract
Here, we demonstrate a powerful method to discriminate DNA mismatches at single-nucleotide resolution from 0 to 5 mismatches (chi0 to chi5) using Kelvin probe force microscopy (KPFM). Using our previously developed method, we quantified the surface potentials (SPs) of individual DNA-capped nanoparticles (DCNPs, approximately 100 nm). On each DCNP, DNA hybridization occurs between approximately 2200 immobilized probe DNA (pDNA) and target DNA with mismatches (tDNA, approximately 80 nM). Thus, each DCNP used in the bioassay (each pDNA-tDNA interaction) corresponds to a single ensemble in which a large number of pDNA-tDNA interactions take place. Moreover, one KPFM image can scan at least dozens of ensembles, which allows statistical analysis (i.e., an ensemble average) of many bioassay cases (ensembles) under the same conditions. We found that as the chin increased from chi0 to chi5 in the tDNA, the average SP of dozens of ensembles (DCNPs) was attenuated owing to fewer hybridization events between the pDNA and the tDNA. Remarkably, the SP attenuation vs. the chin showed an inverse-linear correlation, albeit the equilibrium constant for DNA hybridization exponentially decreased asymptotically as the chin increased. In addition, we observed a cascade reaction at a 100-fold lower concentration of tDNA ( approximately 0.8 nM); the average SP of DCNPs exhibited no significant decrease but rather split into two separate states (no-hybridization vs. full-hybridization). Compared to complementary tDNA (i.e., chi0), the ratio of no-hybridization/full-hybridization within a given set of DCNPs became approximately 1.6 times higher in the presence of tDNA with single mismatches (i.e., chi1). The results imply that our method opens new avenues not only in the research on the DNA hybridization mechanism in the presence of DNA mismatches but also in the development of a robust technology for DNA mismatch detection.
Full Text
http://pubs.rsc.org/en/Content/ArticleLanding/2018/NR/C7NR05250B#!divAbstract
DOI
10.1039/c7nr05250b
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Radiology (영상의학교실) > 1. Journal Papers
Yonsei Authors
Yang, Jae Moon(양재문) ORCID logo https://orcid.org/0000-0001-7365-0395
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/161852
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