Copper-Free Click Chemistry-Mediated Assembly of Single Quantum Dot Nanosensor for Accurately Monitoring Locus-Specific m^(6)A in Cancer Cells  

作　　者：Yue-Ying Li Ning-Ning Zhao Yi-Xuan Geng Qian Han Jian-Ge Qiu Bing-Hua Jiang Zi-Yue Wang Chun-Yang Zhang 

机构地区：[1]Translational Medicine Center,The First Affiliated Hospital of Zhengzhou University,The Academy of Medical Sciences,Henan Institute of Medical and Pharmaceutical Sciences BGI College,Zhengzhou University,Zhengzhou,Henan,450052 China [2]College of Chemistry,Chemical Engineering and Materials Science,Shandong Normal University,Jinan,Shandong,250014 China [3]School of Chemistry and Chemical Engineering,Southeast University,Nanjing,Jiangsu,211189 China

出　　处：《Chinese Journal of Chemistry》2024年第5期499-504,共6页中国化学（英文版）

基　　金：supported by the National Natural Science Foundation of China(Grant No.21735003).

摘　　要：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.

关 键 词：RNA methylation Click chemistry Förster resonance energy transfer BIOSENSOR Cancer 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置] TB383[自动化与计算机技术—控制科学与工程]