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A G-quadruplex DNA-affinity Approach for Purification of Enzymatically Active G4 Resolvase1
Published on: March 18, 2017
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Ratiometric G-quadruplex/hemin DNAzymes with low-dosage associative substrates
Rong Lai1, Xingli Zeng1, Qiuda Xu1
1Key Laboratory of the Ministry of Education for Advanced Catalysis Materials, College of Chemistry and Materials Science, Zhejiang Normal University, Jinhua, 321004, Zhejiang, PR China.
Analytica Chimica Acta
|February 14, 2024
Summary
We developed new associative substrate-based DNAzymes (ASBDs) that use low substrate concentrations and offer a stable colorimetric ratio response. These ASBDs overcome limitations of previous diffusive substrate-based DNAzymes (DSBDs), enabling more robust and environmentally friendly biosensors.
Area of Science:
- Biocatalysis
- Nanotechnology
- Analytical Chemistry
Background:
- Colorimetric G-quadruplex (G4)/hemin DNAzymes are valuable biosensor tools.
- Existing diffusive substrate-based DNAzymes (DSBDs) require high substrate concentrations, produce toxic byproducts, and lack stability.
- There is a need for improved DNAzymes that overcome these limitations.
Purpose of the Study:
- To develop novel associative substrate-based DNAzymes (ASBDs) as an alternative to DSBDs.
- To address the drawbacks of high substrate dosage, product toxicity, and instability in current colorimetric DNAzymes.
- To create more robust, sensitive, and environmentally friendly biosensing platforms.
Main Methods:
- Developed associative substrate-based DNAzymes (ASBDs) utilizing cyanine dyes as associative substrates (ASs).
- Investigated the substrate conversion efficiency and dosage requirements of ASBDs compared to DSBDs.
- Established a colorimetric ratio response for monitoring G4/hemin DNAzyme activity.
- Assessed the stability and tolerance of ASBDs to experimental fluctuations and enzyme cooperation.
Main Results:
- ASBDs required significantly lower substrate dosage (∼10 μM) compared to DSBDs (∼mM).
- Achieved a stable colorimetric ratio response, enhancing tolerance to experimental variations and background noise.
- Demonstrated the ability of ASBDs to work with other enzymes for cascade sensing applications.
- Cyanine dye conversion products exhibited distinct absorption bands, facilitating the ratio response.
Conclusions:
- Developed ASBDs offer a low-dosage, stable colorimetric ratio response, overcoming DSBD limitations.
- ASBDs present a more environmentally benign and low-carbon approach to DNAzyme-based sensing.
- Future design of ASs with diverse color panels can lead to advanced, high-performance sensors.
