化学
费斯特共振能量转移
量子点
核糖核酸
DNA连接酶
荧光
生物物理学
DNA
纳米技术
分子生物学
生物化学
生物
量子力学
基因
物理
材料科学
作者
Yueying Li,Ning‐ning Zhao,Yi‐xuan Geng,Qian Han,Jian‐Ge Qiu,Bing‐Hua Jiang,Ziyue Wang,Chun‐yang Zhang
标识
DOI:10.1002/cjoc.202300595
摘要
Comprehensive Summary N6‐methyladenosine (m 6 A) plays an important role in embryogenesis, nuclear export, transcription splicing, and protein translation control. Herein, we demonstrate a copper‐free click chemistry‐mediated assembly of single quantum dot (QD) nanosensor for accurately monitoring locus‐specific m 6 A in cancer cells. The m 6 A‐sensitive endoribonuclease MazF can digest the unmethylated A‐RNA, and the intact m 6 A‐RNA then hybridizes with DNA probes a and b to produce a sandwich hybrid, initiating the click chemistry to generate probe a–b ligation product via first tandem ligation detection reaction (LDR) cycle. Subsequently, DNA probes c and d can hybridize with the probe a–b ligation product to generate the probe c–d ligation product via second LDR cycle. Both LDR cycles can be repeated through denaturation and annealing reaction to generate abundant biotin‐/fluorophore‐modified probe c–d ligation products that can easily assemble on the QD surface to induce distinct fluorescence resonance energy transfer (FRET) between QD and Cy5. This assay can be homogenously performed without the involvement of copper catalyst, m 6 A‐specific antibody, radioactive labeling, ligase enzyme, enzymatic reverse transcription, and next‐generation sequencing. Moreover, it can discriminate even 0.01% m 6 A level in complex samples and accurately measure cellular m 6 A‐RNA expression, providing a promising avenue for clinical diagnostics and biomedical research.
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