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MicroRNAs (miRNAs) are important cancer-associated biomarkers for early detection. However, accurately measuring miRNAs is challenging due to their low abundance and high homogeneity in human serum. CRISPR technology can greatly improve the specificity and sensitivity of detection in biosensor construction. In this study, we develop a remarkably responsive self-powered biosensing platform for detecting microRNA-21 (miRNA-21) using hybridization chain reaction (HCR) and CRISPR/Cas12a amplification cascade mediated by 3D DNA walkers. Carbon-coated molybdenum disulfide nanotubes with a large surface area and excellent conductivity are fabricated on flexible carbon paper as the anode and cathode for biological fuel cell construction. Glucose oxidase is loaded onto the anode surface, while serving as the biological cathode, it initiated HCR, generating abundant double-stranded DNA. The negatively charged DNA backbone captures electron acceptors [Ru(NH 3) 6 ]3+ through electrostatic adsorption, resulting in a significant open circuit voltage (E OCV ) signal. Through CRISPR/Cas12a amplification cascade mediated by 3D DNA walkers, a smaller E OCV is produced, enabling sensitive target detection. The method exhibits a detection range spanning from 0.0001 to 10,000 pM and capable of detecting as low as 19.0 aM (S/N = 3). This highly selective and sensitive method provides an effective approach for accurately detecting tumor markers in clinical applications. • HCR and 3D DNA Walker-mediated CRISPR/Cas12a cascade amplification strategy is used to fabricate biofuel cell. • Carbon-coated MoS 2 with large surface area and excellent conductivity are fabricated for construction of biofuel cells. • A high-performance self-powered biosensor developed based on biofuel cell for detection of microRNA-21. [ABSTRACT FROM AUTHOR]

MicroRNA-21 (miRNA-21) is widely expressed in various organs and tissues of mammals, including the heart, lungs, brain, pancreas, small intestine, colon, breast, and kidneys, showing high sensitivity and specificity in the diagnosis of multiple types of tumors. It can serve as a new clinical early diagnosis and prognostic biomarker for tumors. This study proposes a self-powered bioplatform, which employs a biological cathode double signal amplification design to construct a high-performance enzyme biofuel cell for the ultra-sensitive detection of tumor biomarker miRNA-21. The biocathodic double signal mechanism involves the catalyzed hairpin self-assembly introduction signal I bilirubin oxidase and a tetrahedral DNA nanostructure that produces a double-helical DNA chain to adsorb signal II electron acceptor [Ru(NH 3) 6 ]3+. This approach not only improves the stability of the sensing system, but also synergistically enhances the open circuit voltage. The developed assay exhibits a wide linear range from 0.1 fM to 1000 pM, with a low detection limit of 0.04 fM (S/N = 3). This study provides a strategy for designing biosensors with improved sensitivity and potential application in medical diagnosis field. • A self-powered biosensor for detection of tumor biomarker is developed using biocathodic double signal amplification design. • CHA and tetrahedral DNA nanostructure can produce a long double-helical DNA chain to introduce signal- electron acceptor. • The developed assay not only improves the stability, but also greatly enhances the sensitivity and selectivity. [ABSTRACT FROM AUTHOR]

Thalassemia is one of the most common monogenic diseases, which seriously affects human growth and development, cardiovascular system, liver, etc. There is currently no effective cure for this disease, making screening for thalassemia particularly important. Herein, a self-powered portable device with high sensitivity and specificity for efficiently screening of low-level thalassemia is developed which is enabled with AuNPs/MoS 2 @C hollow nanorods and triple nucleic acid amplification technologies. It is noteworthy that AuNPs/MoS 2 @C electrode shows the advantages of high electrocatalytic activity, fast carrier migration rate and large specific surface area, which can significantly improve the stability and output signal of the platform. Using high-efficiency tetrahedral DNA as the probe, the target CD122 gene associated with thalassemia triggers a catalytic hairpin assembly reaction to achieve CD122 recycling while providing binding sites for subsequent hybridization chain reaction, greatly improving the detection accuracy and sensitivity of the device. A reliable electrochemical/colorimetric dual-mode assay for CD122 is then established, with a linear response range of 0.0001–100 pM for target concentration and open circuit voltage, and the detection limit is 78.7 aM (S/N = 3); a linear range of 0.0001–10000 pM for CD122 level and RGB Blue value, with a detection limit as low as 58.5 aM (S/N = 3). This method achieves ultra-sensitive and accurate detection of CD122, providing a new method for the rapid and accurate screening of thalassemia. [Display omitted] • Smart AuNPs/Carbon-encased MoS 2 nanorods is prepared to improve the performance of biofuel cell. • A high-efficiency self-powered biosensor is constructed enabled with novel biofuel cell and nucleic acid technique. • A reliable electrochemical/colorimetric dual-mode assay is developed for ultra-sensitive detection of thalassemia. [ABSTRACT FROM AUTHOR]