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Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface

[Image: see text] DNA lesions such as metholcytosine((m)C), 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....

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Autores principales: Wang, Jiajun, Li, Meng-Yin, Yang, Jie, Wang, Ya-Qian, Wu, Xue-Yuan, Huang, Jin, Ying, Yi-Lun, Long, Yi-Tao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2020
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978832/
https://www.ncbi.nlm.nih.gov/pubmed/31989027
http://dx.doi.org/10.1021/acscentsci.9b01129
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author Wang, Jiajun
Li, Meng-Yin
Yang, Jie
Wang, Ya-Qian
Wu, Xue-Yuan
Huang, Jin
Ying, Yi-Lun
Long, Yi-Tao
author_facet Wang, Jiajun
Li, Meng-Yin
Yang, Jie
Wang, Ya-Qian
Wu, Xue-Yuan
Huang, Jin
Ying, Yi-Lun
Long, Yi-Tao
author_sort Wang, Jiajun
collection PubMed
description [Image: see text] DNA lesions such as metholcytosine((m)C), 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 (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 ((m)C, (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.
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spelling pubmed-69788322020-01-27 Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface Wang, Jiajun Li, Meng-Yin Yang, Jie Wang, Ya-Qian Wu, Xue-Yuan Huang, Jin Ying, Yi-Lun Long, Yi-Tao ACS Cent Sci [Image: see text] DNA lesions such as metholcytosine((m)C), 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 (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 ((m)C, (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. American Chemical Society 2020-01-09 2020-01-22 /pmc/articles/PMC6978832/ /pubmed/31989027 http://dx.doi.org/10.1021/acscentsci.9b01129 Text en Copyright © 2020 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Wang, Jiajun
Li, Meng-Yin
Yang, Jie
Wang, Ya-Qian
Wu, Xue-Yuan
Huang, Jin
Ying, Yi-Lun
Long, Yi-Tao
Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title_full Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title_fullStr Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title_full_unstemmed Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title_short Direct Quantification of Damaged Nucleotides in Oligonucleotides Using an Aerolysin Single Molecule Interface
title_sort direct quantification of damaged nucleotides in oligonucleotides using an aerolysin single molecule interface
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978832/
https://www.ncbi.nlm.nih.gov/pubmed/31989027
http://dx.doi.org/10.1021/acscentsci.9b01129
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