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

ABSTRACT 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 Mg 2+ 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.