Rapid and direct detection of m6A methylation by DNAzyme-based and smartphone-assisted electrochemical biosensor

被引：0

作者：

Li, Kai ^{[1
]}

Liang, Yu ^{[1
]}

Li, Xinran ^{[1
]}

Yang, Mengrui ^{[1
]}

Wang, Min ^{[1
]}

Li, Fukai ^{[1
]}

Qi, Xin ^{[1
]}

Zhou, Jian ^{[1
]}

Fu, Wei ^{[2
]}

Li, Liang ^{[1
]}

机构：

[1] Chinese Acad Agr Sci, Inst Qual Stand & Testing Technol Agroprod, Beijing 100081, Peoples R China

[2] Minist Agr & Rural Affairs, Dev Ctr Sci & Technol, Beijing 100176, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2025年 / 267卷

关键词：

DNAzyme; Tetrahedron DNA; Electrochemical biosensor; m(6)A methylation; MESSENGER-RNA; N-6-METHYLADENOSINE RNA;

D O I：

10.1016/j.bios.2024.116788

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

m(6)A methylation detection is crucial for understanding RNA functions, revealing disease mechanisms, guiding drug development and advancing epigenetics research. Nevertheless, high-throughput sequencing and liquid chromatography-based traditional methods still face challenges to rapid and direct detection of m(6)A methylation. Here we report a DNAzyme-based and smartphone-assisted electrochemical biosensor for rapid detection of m(6)A. We initially identified m(6)A methylation-sensitive DNAzyme mutants through site mutation screening. These mutants were then combined with tetrahedral DNA to modify the electrodes, creating a 3D sensing interface. The detection of m(6)A was accomplished by using DNAzyme to capture and cleave the m(6)A sequence. The electrochemical biosensor detected the m(6)A sequence at nanomolar concentrations with a low detection limit of 0.69 nM and a wide detection range from 10 to 10(4) nM within 60 min. As a proof of concept, the 3 '-UTR sequence of rice was selected as the m(6)A analyte. Combined with a smartphone, our biosensor shows good specificity, sensitivity, and easy-to-perform features, which indicates great prospects in the field of RNA modification detection and epigenetic analysis.

引用

页数：10