Multiplexed and Amplified Electronic Sensor for the Detection of MicroRNAs from Cancer Cells

Abstract

The detection of microRNA expression profiles plays an important role in early diagnosis of different cancers. On the basis of the employment of redox labels with distinct potential positions and duplex specific nuclease (DSN)-assisted target recycling signal amplifications, we have developed a multiplexed and convenient electronic sensor for highly sensitive detection of microRNA (miRNA)-141 and miRNA-21. The sensor is constructed by self-assembly of thiol-modified, redox species-labeled hairpin probes on the gold sensing electrode. The hybridizations between the target miRNAs and the surface-immobilized probes lead to the formation of RNA/DNA duplexes, and DSN subsequently cleaves the redox-labeled hairpin probes of the RNA/DNA duplexes to recycle the target miRNAs and to generate significantly amplified current suppression at different potentials for multiplexed detection of miRNA-141 and miRNA-21 down to 4.2 and 3.0 fM, respectively. The sensor is also highly selective toward the target miRNAs and can be employed to monitor miRNAs from human prostate carcinoma (22Rv1) and breast cancer (MCF-7) cell lysates simultaneously. The sensor reported here thus holds great potential for the development of multiplexed, sensitive, selective, and simple sensing platforms for simultaneous detection of a variety of miRNA biomarkers for clinic applications with careful selection of the labels.

MicroRNA-mediated signal amplification coupled with GNP/dendrimers on a mass-sensitive biosensor and its applications in intracellular microRNA quantification

“…Therefore, there are urgent demands for constructing sensitive and selective sensing methods for miRNA detection. Recently, various strategies, such as electrochemistry (Yang et al, 2014), fluorescence (Degliangeli et al, 2014;Wu et al, 2014), chemiluminescence (Deng et al, 2013a(Deng et al, , 2013b, colorimetry (Zhou et al, 2015;Shen et al, 2013), backscattering interferometry (Adams et al, 2013), and mass spectrometry (Popova and Williamson, 2014), etc. have been proposed.…”

Sensitive detection of microRNAs based on the conversion of colorimetric assay into electrochemical analysis with duplex-specific nuclease-assisted signal amplification

“…More importantly, this enzyme can clearly discriminate among mismatched duplexes, even a one-base mismatch. 11,12 Therefore, analytic specificity has been significantly improved with DSN-mediated analytical strategies, including colorimetric assays, [13][14][15] electrochemistry, [16][17][18][19][20][21][22][23][24] fluorescence,…”

“…17,21,22,[53][54][55][56] Furthermore, we demonstrated that the DSN-assisted colorimetric assay can be converted into an electrochemical analysis with improved sensitivity and specificity. Specifically, the DNA probe immobilized on the electrode surface and that labeled with ferrocene (Fc) (denoted "Fc-DNA-Fc") in the solution triggered the in situ assembly of positively charged AuNPs through electrostatic interaction.…”

Coupling hybridization chain reaction with catalytic hairpin assembly enables non-enzymatic and sensitive fluorescent detection of microRNA cancer biomarkers

“…Increasing studies have shown that altered expression profiles of miRNAs are closely associated with a number of diseases, especially human cancers (Lu et al, 2005;Yanaihara et al, 2006;de Planell-Saguer and Rodicio, 2011). Because of their significant biological roles, miRNAs can therefore serve as potential biomarkers for the diagnosis of different cancers (Hammond, 2006;Cissell et al, 2007;Cissell and Deo, 2009;Yang et al, 2014).…”