Visual Detection of Multiplex MicroRNAs Using Cationic Conjugated Polymer Materials

Abstract

A simple, visual, and specific method for simultaneous detection of multiplex microRNAs (miRNAs) has been developed by integrating duplex-specific nuclease (DSN)-induced amplification with cationic conjugated polymer (CCP) materials. The probe DNA with a complementary sequence to target miRNA is labeled with fluorescein dye (FAM). Without target miRNA, the single-strand DNA probe cannot be digested by DSN. Upon adding CCPs, efficient fluorescence resonance energy transfer (FRET) from CCP to FAM occurs owing to strong electrostatic interactions between CCP and the DNA probe. In the presence of target miRNA, the DNA probe hybridizes with target miRNA followed by digestion to small nucleotide fragments by DSN; meanwhile, the miRNA is released and subsequently interacts again with the probe, resulting in the cycled digestion of the DNA probe. In this case, weak electrostatic interactions between oligonucleotide fragments and CCP lead to inefficient FRET from CCP to FAM. Thus, by triggering the FRET signal from CCP to FAM, miRNA can be specially detected, and the fluorescence color change based on FRET can be visualized directly with the naked eye under an UV lamp. Furthermore, an energy transfer cascade can be designed using CCP and DNA probes labeled at the 5'-terminus with FAM and Cy3 dyes, and the multistep FRET processes offer the ability of simultaneous detection of multiplex miRNAs.

MicroRNA Quantitation During Dendritic Cell Endocytosis Using Imaging Flow Cytometry: Key Factors and Requirements

“…6). In this system, the only method for detecting miR-590-5p-Cy3 fluorescence is to combine it with anti-miR-590-5p-Fam [35]. As expected, under an excitation of 200 mW at 488 nm, few cells were found in the "high intensity" region of the Cy3 channel (Fig.…”

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

“…[72] Figure 11 The LAMP assay for miRNA detection [72] . [79] Figure 12 The miRNA detection based on DSN and CCP [79] . 质适用于 miRNA 的检测 [74][75][76][77] [81] Figure 13 MB probe and T7 Exo-assisted cyclic amplification for miRNA detection [81] .…”

Dimedone‐decorated conjugated polymer: Tandem knoevenagel‐michael post‐modification synthesis and its application as optical probe for Hg2+ and ClO− in high‐water‐fraction mediums

“…Fluorescent conjugated polymers(CPs)‐based optical probes, highlighted for their high sensitivity and selectivity, have been regarded as promising platforms for monitoring specific ions and biological targets . Up to date, various fluorescent CPs have been developed for detecting poisonous ions, explosive, thiols, and mRNAs . The common designing principle for these probes is based on the incorporation of reacting/binding sites, which can specifically interact with detecting targets, into the chemical structures of fluorescent CPs .…”