Chemical Engineering of DNAzyme for Effective Biosensing and Gene Therapy

Abstract RNA‐cleaving DNAzymes are in vitro selected functional nucleic acids with inherent catalytic activities. Due to their unique properties, such as high specificity, substrate cleavage capability, and programmability, DNAzymes have emerged as powerful tools in the fields of analytical chemistry, chemical biology, and biomedicine. Nevertheless, the biological applications of DNAzymes are still impeded by several challenges, such as structural instability, compromised catalytic activity in biological environments and the lack of spatiotemporal control designs, which may result in false‐positive signals, limited efficacy or non‐specific activation associated with side effects. To address these challenges, various strategies have been explored to regulate DNAzyme activity through chemical modifications, enhancing their stability, selectivity, and functionality, thereby positioning them as ideal candidates for biological applications. In this review, a comprehensive overview of chemically modified DNAzymes is provided, discussing modification strategies and the effects of these modifications on DNAzymes. Specific examples of the use of chemically modified DNAzymes in biosensing and gene therapy are also presented and discussed. Finally, the current challenges in the field are addressed and offer perspectives on the potential direction for chemically modified DNAzymes.