Duplex-specific nuclease-mediated bioanalysis

•Duplex-specific nucleases (DSNs) can selectively cleave DNA within RNA/DNA heteroduplexes. •We review the applications of DSNs in biological and biomedicine fields. •DSNs hold promise for the development of amplification-free methods for microRNA detection. •Challenges and perspectives of DSNs are discussed. Duplex-specific nucleases (DSNs) are promising tools for bioanalysis because of their unique ability to cleave DNA within duplexes while keeping a single strand intact. There is prevalent use of DSNs in both biomedical and biological science applications, such as cDNA library construction, circulating miRNA detection, telomeric overhang detection, and SNP recognition. We present an overview of the current knowledge of DSNs, with special emphasis on DSN-mediated isothermal signal amplification strategies for trace miRNA detection. Continued innovation to address key challenges, such as amplification-free approaches, will open up new avenues in the field of miRNA profiling, offering opportunities for improved personalized medicine, preventive medicine, and translational medicine. Duplex-specific nucleases (DSNs) are promising tools for bioanalysis because of their unique ability to cleave DNA within duplexes while keeping a single strand intact. There is prevalent use of DSNs in both biomedical and biological science applications, such as cDNA library construction, circulating miRNA detection, telomeric overhang detection, and SNP recognition. We present an overview of the current knowledge of DSNs, with special emphasis on DSN-mediated isothermal signal amplification strategies for trace miRNA detection. Continued innovation to address key challenges, such as amplification-free approaches, will open up new avenues in the field of miRNA profiling, offering opportunities for improved personalized medicine, preventive medicine, and translational medicine.