Mutation detection by electro catalysis at DNA-modified electrodes

Elizabeth M. Boon; Donato M. Ceres; Thomas G. Drummond; Michael G. Hill; Jacqueline K. Barton

doi:10.1038/80301

Mutation detection by electro catalysis at DNA-modified electrodes

Elizabeth M. Boon
, Donato M. Ceres
, Thomas G. Drummond
, Michael G. Hill
, Jacqueline K. Barton

Chemistry

California Institute of Technology
Occidental College

Research output: Contribution to journal › Article › peer-review

698 Scopus citations

Abstract

Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)₆]^3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and "hot spot" mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection.

Original language	English
Pages (from-to)	1096-1100
Number of pages	5
Journal	Nature Biotechnology
Volume	18
Issue number	10 SUPPL.
DOIs	https://doi.org/10.1038/80301
State	Published - Oct 1 2000

Keywords

Damage detection
DNA chip
DNA monolayer
DNA-mediated electron transfer
Electrocatalysis
Electrochemistry
Methylene blue
Mutation detection
P53 gene
SNPs

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10.1038/80301

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@article{6eb8709efbe649dcad61dfb5d005efde,

title = "Mutation detection by electro catalysis at DNA-modified electrodes",

abstract = "Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)6]3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and {"}hot spot{"} mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection.",

keywords = "Damage detection, DNA chip, DNA monolayer, DNA-mediated electron transfer, Electrocatalysis, Electrochemistry, Methylene blue, Mutation detection, P53 gene, SNPs",

author = "Boon, \{Elizabeth M.\} and Ceres, \{Donato M.\} and Drummond, \{Thomas G.\} and Hill, \{Michael G.\} and Barton, \{Jacqueline K.\}",

year = "2000",

month = oct,

day = "1",

doi = "10.1038/80301",

language = "English",

volume = "18",

pages = "1096--1100",

journal = "Nature Biotechnology",

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T1 - Mutation detection by electro catalysis at DNA-modified electrodes

AU - Boon, Elizabeth M.

AU - Ceres, Donato M.

AU - Drummond, Thomas G.

AU - Hill, Michael G.

AU - Barton, Jacqueline K.

PY - 2000/10/1

Y1 - 2000/10/1

N2 - Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)6]3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and "hot spot" mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection.

AB - Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)6]3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and "hot spot" mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection.

KW - Damage detection

KW - DNA chip

KW - DNA monolayer

KW - DNA-mediated electron transfer

KW - Electrocatalysis

KW - Electrochemistry

KW - Methylene blue

KW - Mutation detection

KW - P53 gene

KW - SNPs

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U2 - 10.1038/80301

DO - 10.1038/80301

M3 - Article

C2 - 11017050

AN - SCOPUS:0033772184

SN - 1087-0156

VL - 18

SP - 1096

EP - 1100

JO - Nature Biotechnology

JF - Nature Biotechnology

IS - 10 SUPPL.

ER -

Mutation detection by electro catalysis at DNA-modified electrodes

Abstract

Keywords

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