Aims: To establish a FF1-ATP synthase molecular motor biosensor to accurately identify colon cancer miRNAs.
Main Methods: The FF1-ATP synthase molecular motor is extracted by fragmentation-centrifugation and connected to the colon cancer-specific miR-17 capture probe in the manner of the ε subunit-biotin-streptavidin-biotin system. Signal probes are designed for dual-signal characterization to increase detection accuracy. The FF1-ATPase rotation rate decreases when the signaling and capture probes are combined with the target miRNA, resulting in a decrease in ATP synthesis. miR-17 concentrations are determined by changes in ATP-mediated chemiluminescence intensity and signal probe-mediated OD.
Key Findings: The chemiluminescence intensity and OD show a good linear relationship with the miR-17 concentration in the range of 5 to 200 nmol L (R = 0.9985, 0.9989). The colon cancer mouse model is established for the blood samples, and miR-17 in serum and RNA extracts is quantitatively determined using the constructed sensor.
Significance: The results are consistent with colon cancer progression, and the low concentration of miR-17 detecting accuracy is comparable to the PCR assay. In conclusion, the developed method is a direct, rapid, and promising method for miRNA detection of colon cancer.
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http://dx.doi.org/10.1016/j.lfs.2023.121527 | DOI Listing |
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