A fluorescence strategy for the rapid detection of miRNA-21 based on G-quadruplex and cyclic amplification signal

Ying Yan; Tingting Hu; Yukang Fang; Xinying Xiang; Changbei Ma

doi:10.1016/j.ab.2022.114775

A fluorescence strategy for the rapid detection of miRNA-21 based on G-quadruplex and cyclic amplification signal

Anal Biochem. 2022 Sep 1:652:114775. doi: 10.1016/j.ab.2022.114775. Epub 2022 Jun 3.

Authors

Ying Yan¹, Tingting Hu¹, Yukang Fang¹, Xinying Xiang¹, Changbei Ma²

Affiliations

¹ School of Life Sciences, Central South University, Changsha, 410013, China.
² School of Life Sciences, Central South University, Changsha, 410013, China. Electronic address: macb2012@csu.edu.cn.

PMID: 35667449
DOI: 10.1016/j.ab.2022.114775

Abstract

MicroRNAs (miRNAs) are commonly used as biomarkers for the diagnosis of tumors. Since miRNA expression is strongly correlated to carcinogenesis, the detection of miRNA concentration in cells would be valuable for the diagnosis and evaluation of tumors. In this study, we proposed a system using two strands of DNA, one modified by a phosphate group at the 5' end, called Cap, and the other with a hairpin structure, called HP. The Cap chain could open the hairpin structure of HP and expose the sequences rich in G bases to form the G-quadruplex structure. Then, a strong fluorescence signal was emitted in the presence of N-methyl mesoporphyrin IX (NMM). However, with the addition of miRNA-21, Cap hybridized with it to form double chains, which were then cleaved by the digestion of lambda exonuclease, resulting in a weak fluorescent signal. The proposed method could detect miRNA-21 at a concentration of 1.4 pM with a broad dynamic linear range from 5 pM to 5 nM.

Keywords: G-quadruplex; Hairpin probe; Lambda exonuclease; NMM; miRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biosensing Techniques* / methods
DNA / chemistry
G-Quadruplexes*
Limit of Detection
MicroRNAs*
Spectrometry, Fluorescence

Substances

MicroRNAs
DNA