Advances in DNAzyme Selection, Molecular Engineering and Biomedical Applications

Area of Science:

• Biochemistry and Molecular Biology
• Synthetic Biology
• Nanotechnology

Background:

• DNAzymes are catalytic DNA molecules with potential in various applications.
• Current DNAzyme performance in biological settings faces limitations.
• A need exists for improved design and application strategies for DNAzymes.

Purpose of the Study:

• To establish a novel conceptual framework for DNAzyme development.
• To integrate selection, engineering, and application into a unified workflow.
• To enhance DNAzyme performance for biological and clinical use.

Main Methods:

• Evolution of SELEX (Systematic Evolution of Ligands by Exponential Enrichment) with application-driven selection.
• Incorporation of functional/environmental constraints and deep sequencing for high-throughput readouts.
• Molecular engineering strategies including 2'-sugar modifications, XNA substitution, and scaffold-based organization.
• Structure- and computation-guided design approaches.

Main Results:

• Demonstrated a continuous iteration workflow for DNAzyme development.
• Summarized engineering strategies to improve DNAzyme stability, kinetics, and controllability.
• Surveyed diverse applications including biosensing, imaging, and therapeutics.
• Outlined translational priorities for clinical deployment.

Conclusions:

• The proposed framework enhances DNAzyme development by unifying selection, engineering, and application.
• Improved DNAzymes show promise for ultrasensitive diagnostics, advanced imaging, and targeted therapies.
• Translational priorities focus on data-driven design, delivery, safety evaluation, and scalable manufacturing for clinical use.