Journal
ACS SENSORS
Volume -, Issue -, Pages -Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.1c02748
Keywords
biomarkers; miRNA; fluorescence resonance energy transfer (FRET); high-confidence; single-molecule; triple negative breast cancer (TNBC)
Funding
- National Center for Advancing Translational Sciences [UL1TR002649]
- CCTR Endowment Fund of Virginia Commonwealth University
- National Institutes of Health/National Cancer Institute [CCR19608826]
- Susan G. Komen Foundation [1R01CA246182]
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In this study, a DNA-based FRET sensor was developed for ultrasensitive detection of a miRNA biomarker expressed by TNBC cells. The sensor showed high specificity, low detection limit, and a large dynamic range, making it suitable for detecting different miRNAs and enabling early diagnosis and screening of other genetic disorders.
MicroRNAs (miRNAs) play a crucial role in regulating gene expression and have been linked to many diseases. Therefore, sensitive and accurate detection of disease-linked miRNAs is vital to the emerging revolution in early diagnosis of diseases. While the detection of miRNAs is a challenge due to their intrinsic properties such as small size, high sequence similarity among miRNAs and low abundance in biological fluids, the majority of mi RNA-detection strategies involve either target/signal amplification or involve complex sensing designs. In this study, we have developed and tested a DNA-based fluorescence resonance energy transfer (FRET) sensor that enables ultrasensitive detection of a miRNA biomarker (miRNA-342-3p) expressed by triple-negative breast cancer (TNBC) cells. The sensor shows a relatively low FRET state in the absence of a target but it undergoes continuous FRET transitions between low-and high-FRET states in the presence of the target. The sensor is highly specific, has a detection limit down to low femtomolar (fM) without having to amplify the target, and has a large dynamic range (3 orders of magnitude) extending to 300 000 fM. Using this strategy, we demonstrated that the sensor allows detection of miRNA-342-3p in the miRNA-extracts from cancer cell lines and TNBC patient-derived xenografts. Given the simple to-design hybridization-based detection, the sensing platform developed here can be used to detect a wide range of miRNAs enabling early diagnosis and screening of other genetic disorders.
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