Enzyme-free and multiplexed micro RNA detection using micro RNA-initiated DNA molecular motor

作     者：hui wang honghong wang chenghui liu xinrui duan zhengping li 

作者机构：Key Laboratory of Applied Surface and Colloid ChemistryMinistry of EducationKey Laboratory of Analytical Chemistry for Life Science of Shaanxi ProvinceSchool of Chemistry and Chemical EngineeringShaanxi Normal UniversityXi'an 710062China 

出 版 物：《Science China Chemistry》 (中国科学（化学英文版）)

年 卷 期：2016年第59卷第1期

页      面：83-88页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 

基　　金：the National Natural Science Foundation of China(21335005,21472120) the Fundamental Research Funds for the Central Universities(GK201501003,GK201303003) the Excellent Doctor Innovation Project of Shaanxi Normal University 

主　　题：multiplexed micro RNA detection DNA molecular motor enzyme free synchronous fluorescence spectrum 

摘      要：In this work,we have developed a sensitive,simple,and enzyme-free assay for detection of micro RNAs(mi RNAs)by means of a DNA molecular motor consisting of two stem-loop DNAs with identical stems and complementary loop *** the presence of mi RNA target,it can hybridize with one of the stem-loop DNA to open the stem and to produce a mi RNA/DNA hybrid and a single strand(ss)DNA,the ss DNA will in turn hybridize with another stem-loop DNA and finally form a double strand(ds)DNA to release the mi *** of the stem-loop DNA is double-labeled by a fluorophore/quencher pair with efficiently quenched *** formation of ds DNA can produced specific fluorescence signal for mi RNA *** released mi RNA will continuously initiate the next hybridization of the two stem-loop DNAs to form a cycle-running DNA molecular motor,which results in great fluorescence *** the efficient signal amplification,as low as 1 pmol/L mi RNA target can be detected and a wide dynamic range from 1 pmol/L to 2 nmol/L is also ***,by designing different stem-loop DNAs specific to different mi RNA targets and labeling them with different fluorophores,multiplexed mi RNAs can be simultaneously detected in one-tube reaction with the synchronous fluorescence spectrum(SFS)technique.