<br/>Hybridization probes have been used in the detection of specific nucleic acids for the last 50 years. Current challenges of the technology include (1) detection of single nucleotide variations (SNV) under ambient conditions (2) tight binding to folded nucleic acids, and (3) the cost of fluorescent probes. Here we introduce a sensor (called OWL2), which addresses all these issues. The sensor uses two analyte binding arms to tightly bind and unwind folded nucleic acid, two sequence-specific strands to recognize the analyte sequence with high selectivity, and a universal molecular beacon (UMB) probe to signal the recognition event. OWL 2 was able to differential SNV in folded DNA analyte in the range 5-40 oC. The design is cost-efficient since the same fluorescently labeled UMB probe can be used for the detection of analyte sequence.

OWL2: A molecular beacon -based hybridization sensor for highly selective SNP detection of folded nucleic acids

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ACS Fall 2022

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