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Published on: December 23, 2022
Sensitive Small Molecule Aptasensing based on Hybridization Chain Reaction and CRISPR/Cas12a Using a Portable
Long Ma1, Dan Liao1, Zhiying Zhao1
1State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China.
Researchers developed SMART-Cas12a, a novel CRISPR/Cas biosensing platform for detecting non-nucleic acid small molecules. This adaptable method offers sensitive, specific, and on-site detection, expanding CRISPR applications.
Area of Science:
- Biotechnology
- Molecular Biology
- CRISPR/Cas Systems
Background:
- CRISPR/Cas biosensors excel at nucleic acid detection.
- Expanding CRISPR/Cas systems for non-nucleic acid targets is crucial for broader applications.
- Current methods for small-molecule detection using CRISPR/Cas are limited.
Purpose of the Study:
- To develop a versatile biosensing platform for non-nucleic acid small-molecule detection using CRISPR/Cas12a.
- To establish an efficient signal amplification strategy for small-molecule targets.
- To enable rapid, on-site detection with smartphone integration.
Main Methods:
- Developed SMART-Cas12a (small-molecule aptamer regulated test using CRISPR/Cas12a).
- Utilized hybridization chain reaction for signal amplification triggered by aptamer-target binding.
- Integrated CRISPR/Cas12a for amplified product recognition and trans-cleavage activation.
- Employed a 3D-printed visualizer and smartphone app for signal readout.
Main Results:
- Demonstrated detection of adenosine triphosphate (ATP) as a model target.
- Achieved a linear detection range from 0.1 to 750 μM for ATP.
- Obtained a low detection limit of 1.0 nM for ATP.
- Showcased satisfactory selectivity and recovery rates in complex samples.
- Completed sample-to-answer analysis in under 100 minutes.
Conclusions:
- SMART-Cas12a expands CRISPR/Cas applications to non-nucleic acid small-molecule detection.
- The platform offers high adaptability, sensitivity, specificity, and on-site capabilities.
- This work provides a generalized prototype for detecting diverse analytes using CRISPR/Cas technology.
