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Protein-Controlled Split DNAzyme to Enhance Catalytic Activity: Design and Performance.
Lingying Xia1,2, Lijie Du1, Xiandeng Hou1
1Analytical & Testing Center, Sichuan University, Sichuan, Chengdu 610064, PR China.
Analytical Chemistry
|July 16, 2024
Summary
Protein control enhances split DNAzyme (Pc SD) catalytic activity by increasing local fragment concentration and improving cofactor binding. This innovation offers sensitive diagnostic tools for biomarkers like AFP.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Split DNAzymes offer catalytic activity but can suffer from low efficiency.
- Controlling DNAzyme assembly is crucial for enhancing catalytic performance.
Purpose of the Study:
- To develop protein-controlled split DNAzymes (Pc SD) with enhanced catalytic activity.
- To investigate the impact of protein-induced assembly on DNAzyme conformation and cofactor binding.
- To explore the influence of split site positioning on Pc SD activity and its application in diagnostics.
Main Methods:
- Utilized proteins and affinity ligands to control the assembly of split DNAzyme fragments.
- Systematically varied split sites within the catalytic core of DNAzymes.
- Assessed catalytic activity and cleavage efficiency of Pc SD compared to free split DNAzymes.
- Developed diagnostic assays using Pc SD for streptavidin and AFP biomarker detection.
Main Results:
- Protein control significantly increased the local concentration of split DNAzyme fragments, promoting reassembly.
- Pc SD exhibited enhanced cleavage efficiency and cofactor affinity compared to free split DNAzymes.
- Optimal split site positioning was identified, influencing Pc SD activity.
- Developed sensitive diagnostic tools with low limits of detection (LOD) for streptavidin (0.1 pM) and AFP (2 pM in human serum).
Conclusions:
- Protein-mediated control is an effective strategy for boosting DNAzyme catalytic activity.
- Pc SD technology provides a robust platform for developing sensitive and rapid diagnostic tools.
- This work offers fundamental insights into optimizing DNAzyme catalysis for various applications.
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