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Photoinduced Electron Transfer‐Based Fluorescence Quenching Combined with Rolling Circle Amplification for Sensitive Detection of MicroRNA
Author(s) -
Zhou Fulin,
Meng Rong,
Liu Qiang,
Jin Yan,
Li Baoxin
Publication year - 2016
Publication title -
chemistryselect
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.437
H-Index - 34
ISSN - 2365-6549
DOI - 10.1002/slct.201601485
Subject(s) - fluorescence , quenching (fluorescence) , rolling circle replication , photoinduced electron transfer , microrna , photochemistry , chemistry , electron transfer , materials science , biophysics , biology , physics , optics , gene , biochemistry , dna replication
MicroRNAs (miRNAs) detection is crucial for further understanding the biological functions of miRNAs and early cancer diagnosis and therapeutics, but now remains a great challenge. Here, we present a novel homogenous biosensing strategy for fluorescence detection of miRNA. In this strategy, a preliminary synthesized circular DNA was used as recognition probe for hybridization with miRNA target, and then miRNA target primed a rolling circle amplification (RCA) reaction. The RCA product could be hybridized with thousands of carboxyfluorescein (FAM)‐labeled linear DNA probes, which led FAM to be close to ‐GGG‐ base of RCA product, accompanying with the significant fluorescence quenching due to photoinduced electron transfer (PET) between FAM and guanine. This is for the first time to integrate RCA and PET into one detection process. With highly efficient amplification of RCA and excellent signal readout of PET, this method exhibited a high sensitivity toward target miRNA with a detection limit of 6 aM. The target‐dependent circularization of the padlock probe and the ligation reaction could improve the specificity effectively, leading to discrimination between miRNA family members. This method provides a simple, isothermal, and low‐cost approach for sensitive detection of miRNA and holds great potential for early diagnosis in gene‐related diseases.