Amplified Tandem Spinach-Based Aptamer Transcription Enables Low Background miRNA Detection

Analytical Chemistry
Xin TangJinghong Li

Abstract

MicroRNAs (miRNAs) play key roles in regulating gene expression and cell functions, which are recognized as potential biomarkers for many human diseases. Sensitive, specific, and reliable detection of miRNA is highly demanded for clinical diagnosis and therapy. Herein, we describe a label-free and low-background fluorescent assay, termed amplified tandem Spinach-based aptamer transcription assay (AmptSpi assay) for highly sensitive miRNA detection by polymeric rolling circle amplicon mediated multiple transcription. Target miRNA is recognized by padlock probe to form polymeric rolling circle amplicon. Then the following transcription process rapidly produces large amounts of repeats of RNA Spinach aptamers, lightened up by the addition of fluorescent dye DFHBI for miRNA quantitative analysis, achieving label-free and nearly zero-background. Besides, the assay could also confer high selectivity to distinguish miRNA among the miRNA family members with 1- or 2-nucleotide (nt) difference. This method was capable of completing detection in human serum sample or cell extracts in hours, indicating great potential in the early diagnosis of diseases.

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Citations

Jan 19, 2020·Chemistry : a European Journal·Li PanYi-Ming Liu
Mar 17, 2020·Chembiochem : a European Journal of Chemical Biology·Jiuxing LiYingfu Li
Nov 16, 2019·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yiting WangJianhua Wang
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Nov 5, 2019·Frontiers in Pharmacology·Kyung-Ho Lee, Dong-Myung Kim
Jan 19, 2021·Chemical Communications : Chem Comm·Jisu WooKi Soo Park
Mar 6, 2021·Chemical Reviews·Lingling WuWeihong Tan
Aug 14, 2019·Analytical Chemistry·Mingshu XiaoHao Pei
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May 1, 2019·ACS Synthetic Biology·Shue WangAllen P Liu
Jan 27, 2022·Chemical Communications : Chem Comm·Yoonseo HongYong-Beom Shin

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