Electrochemical immunosensor for N6-methyladenosine RNA modification detection

•An electrochemical immunosensor was fabricated for RNA methylation detection.•N6-methyladenosine antibody was used as capture probe for RNA methylation.•The developed method showed wide linear range from 0.01 to 10nM and the detection limit was 2.57pM.•The immunosensor was applied to detect the met...

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Bibliographic Details
Published in:Sensors and actuators. B, Chemical Chemical, 2015-12, Vol.221, p.1-6
Main Authors: Yin, Huanshun, Zhou, Yunlei, Yang, Zhiqing, Guo, Yunlong, Wang, Xinxu, Ai, Shiyun, Zhang, Xiansheng
Format: Article
Language:English
Subjects:
Anti-N6-methyladenosine antibody
Antibodies
Biosensors
Cytokinin
Electrodes
Gene expression
Glassy carbon
Immunosensor
Immunosensors
Methylation
mRNA
N6-methyladenosine
Ribonucleic acids
RNA methylation
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Summary:•An electrochemical immunosensor was fabricated for RNA methylation detection.•N6-methyladenosine antibody was used as capture probe for RNA methylation.•The developed method showed wide linear range from 0.01 to 10nM and the detection limit was 2.57pM.•The immunosensor was applied to detect the methylation level of mRNA in rice seedling leaves. N6-methyladenosine (m6A) is a ubiquitous modification in mRNA and other RNAs across most eukaryotes, which is likely dynamically regulated throughout developmental processes. In this work, an electrochemical immunosensor was fabricated for RNA methylation detection using gold nanoparticles modified glassy carbon electrode (AuNPs/GCE) as substrate electrode. The assembly of 4-aminophenylboronic acid on AuNPs/GCE triggered the further immobilization of anti-m6A antibody on the substrate electrode through the specific interaction between boric acid and glycosyl of antibody, which led to the following capture of methylated RNA through the immunoreaction. A decreased electrochemical reduction signal of the redox probe of [Fe(CN)6]3− was obtained based on the synergistic effect of electrostatic repulsion and steric hindrance. The developed method showed a wide linear range for methylated RNA from 0.01 to 10nM and the detection limit was 2.57pM (S/N=3). Moreover, the down-regulated expression of methylation level of mRNA in the leaves of rice seedlings after incubation with 6-benzylaminopurine was confirmed by the fabricated biosensor, indicating the great potential for assaying the expression level of methylated RNA in real sample.
ISSN:0925-4005
1873-3077
DOI:10.1016/j.snb.2015.06.045