The 10-23 DNAzyme in Biosensing and Diagnostics: Applications, Challenges, and Future Directions

Catalytic DNA molecules (DNAzymes) have garnered increasing attention as components of biosensing and diagnostic platforms due to their simplicity, programmability, and cost-effectiveness. Among them, the 10-23 DNAzyme remains the most widely used RNA-cleaving DNAzyme, combining high catalytic efficiency with broad adaptability across diverse sensor architectures. Despite these advantages, its performance can be significantly hindered by suboptimal reaction temperature, low Mg²⁺ concentrations, nuclease-rich biological matrices, and restricted accessibility to structured RNA targets. Such limitations have impeded its widespread adoption in simple, robust point-of-care formats. This review examines the integration of the 10-23 DNAzyme into contemporary biosensing and diagnostic systems-including colorimetric, fluorescent, electrochemical, electrochemiluminescent, and intracellular sensors-highlighting both direct and regulated activation strategies and the dual role of 10-23 as a molecular recognition element and signal reporter. We also discuss key challenges in catalytic performance, stability, assay workflow, and clinical validation, as well as emerging solutions such as chemical modifications, nanoparticle-based protection, and advanced sensor architectures. Together, these insights outline the current landscape and future opportunities for advancing the 10-23 DNAzyme toward next generation biosensing and diagnostic applications.