Enhancing the miRNA Detection Sensitivity of DNA Origami Book Biosensors Using Lock Modifications and a Polymer Additive
Year: 2025
Type: article
Abstract: DNA origami-based biosensors provide a powerful and programmable platform for sensitive and specific molecular diagnostics. In this study, a dynamic DNA origami book biosensor is validated for direct detection of microRNA21 (miR-21) in complex biological fluids, including human serum and plasma. Two optical readouts are engineered and tested: Förster resonance energy transfer (FRET) and fluorescence quenching using precisely arranged donor-acceptor fluorophore pairs. To improve sensitivity, detection occurs under varied conditions, including modifications to the lock mechanism (fluorine-modified bases and locked nucleic acid) and the addition of a biocompatible polymer, diethylene glycol (DEG). The biosensor maintains specificity in spiked serum, plasma, and clinical plasma samples from breast cancer patients. Single origami nanostructure detection experiments show that target miRNAs are identified within 10 min, with limits of detection (LoD) of 0.69 pm in buffer with LNA, 8.86 pm in 100% serum, and 23.24 pm in 100% plasma, without enzymatic amplification. Importantly, simultaneous detection of miR-21 and miR-7a in clinical plasma samples is demonstrated, underscoring the platform's potential for personalized diagnostics and liquid biopsy applications. These results show that tuning the lock mechanism and adding polymers enhances sensitivity and enables adaptable performance, advancing nanotechnology-based biomarker profiling, including diverse microRNA detection.
Source: Small
Institution: University of Fribourg
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,790
Funders: National Science Foundation, European Commission, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, Innosuisse - Schweizerische Agentur für Innovationsförderung, National Center of Competence in Research Bio-Inspired Materials, University of Fribourg, HORIZON EUROPE European Research Council
Awards: G2535156004, microRNA assay system based on self-assembled nanoscale DNA origami arrays, G434337789, DNA self-assembled optical nano-antennas for single photon emitters. Directing and concentrating light for future diagnostic platforms, NCCR Bio-Inspired Materials: Using Concepts from Nature to Create ‚Smart' Materials (phase II), G7686874183, G894609541, G993971353
OpenAlex ID: W4416226765