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Fusing Allosteric Ribozymes with CRISPR-Cas12a for Efficient Diagnostics of Small Molecule Targets
Lichuan Guo1,2, Shu Zhang1, Xinyu Du1,3
1Key Laboratory of Breast Cancer in Shanghai, Department of Breast Surgery and Department of gynecologic oncology, Institutes of Biomedical Sciences, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
Small Methods
|October 18, 2024
Summary
This study presents a new CRISPR-Cas12a diagnostic system for detecting non-nucleic-acid targets. The system uses allosteric ribozymes for rapid, accurate analysis of small molecules in blood samples.
Area of Science:
- Biotechnology
- Molecular Biology
- Diagnostics
Background:
- CRISPR-Cas systems are versatile molecular tools for genetic manipulation and diagnostics.
- Current CRISPR-Cas diagnostic methods are limited for non-nucleic-acid targets.
Purpose of the Study:
- To develop a novel CRISPR-Cas system for efficient quantitative analysis of non-nucleic-acid targets.
- To demonstrate the system's applicability in real-world samples like blood.
Main Methods:
- Fusion of ligand-dependent allosteric ribozymes with CRISPR-Cas12a.
- Quantitative analysis of small molecules using the developed CRISPR-Cas12a system.
- Adaptation of the system using allosteric deoxyribozymes for enhanced stability.
Main Results:
- The developed system enables quantitative analysis of non-nucleic-acid targets within 1-2 hours.
- Demonstrated generality, reliability, and accuracy on two different small molecules.
- Showcased high-throughput detection capability for small molecules in blood samples.
Conclusions:
- The (deoxy)ribozyme-assisted CRISPR-Cas12a system offers efficient bioanalysis for non-nucleic-acid targets.
- The system's adaptability and performance suggest potential for clinical translation.
- This advancement broadens the application scope of CRISPR-Cas technology in diagnostics.
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