Detection of locus-specific -methyladenosine modification based on Ag-assisted ligation and supersandwich signal amplification
Emerging evidence reveals that the epitranscriptomic mark N 6 -methyladenosine (m 6 A) plays vital roles in organisms, including gene regulation and disease progression. However, developing sensitive methods to detect m 6 A modification, especially the identification of m 6 A marks at the single-sit...
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Published in | Analyst (London) Vol. 146; no. 4; pp. 1355 - 136 |
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Main Authors | , , , , , , |
Format | Journal Article |
Published |
22.02.2021
|
Online Access | Get full text |
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Summary: | Emerging evidence reveals that the epitranscriptomic mark
N
6
-methyladenosine (m
6
A) plays vital roles in organisms, including gene regulation and disease progression. However, developing sensitive methods to detect m
6
A modification, especially the identification of m
6
A marks at the single-site level, remains a challenge. Therefore, based on target-specific triggered signal amplification, we developed a highly sensitive electrochemical method to detect site-specific m
6
A modifications in DNA. In this work, the m
6
A site in DNA can restrict the ligation assisted by Ag
+
, and this restriction effect can activate the subsequent strand displacement reaction and hybridization chain reaction (HCR), thus achieving signal amplification from the m
6
A site, and finally realizing high sensitivity analysis of m
6
A methylation. Benefiting from the high specificity of base pairs and the extremely weak binding affinity between Ag
+
and m
6
A, the proposed method was used for not only detecting the target DNA with a putative m
6
A site, but also identifying m
6
A marks at the single-site level in DNA. In addition, this study does not rely on antibodies and radiolabeling, so it has the advantage of cost-effectiveness. Therefore, we believe that the proposed strategy may provide a new perspective for methylation research, which can be used to test more clinical samples in further research.
Emerging evidence reveals that the epitranscriptomic mark
N
6
-methyladenosine (m
6
A) plays vital roles in organisms, including gene regulation and disease progression. |
---|---|
Bibliography: | Electronic supplementary information (ESI) available. See DOI 10.1039/d0an02214d |
ISSN: | 0003-2654 1364-5528 |
DOI: | 10.1039/d0an02214d |