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CRISPR-Enhanced Hydrogel Microparticles for Multiplexed Detection of Nucleic Acids
Yoon Ho Roh1,2, Chang Yeol Lee1,3, Sujin Lee1,2
1Institute for Basic Science (IBS), Center for Nanomedicine, Seoul, 03722, Republic of Korea.
Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|February 1, 2023
Summary
Researchers developed CLAMP, a novel hydrogel-based CRISPR/Cas12 system. This method enables fast, sensitive, and multiplexed DNA detection, even in clinical samples like human papillomavirus.
Area of Science:
- Molecular Biology
- Biotechnology
- Analytical Chemistry
Background:
- CRISPR/Cas systems provide sequence-specific detection but multiplexing is challenging.
- Existing multiplexed CRISPR/Cas assays often require complex methodologies.
Purpose of the Study:
- To develop a streamlined and multiplexed CRISPR/Cas12 assay.
- To enable sensitive and rapid detection of nucleic acid targets.
Main Methods:
- Developed CLAMP (Cas-Loaded Annotated Micro-Particles), a hydrogel-based CRISPR/Cas12 system.
- Utilized spatially-encoded hydrogel microparticles (HMPs) for reaction compartmentalization.
- Integrated HMPs with a microfluidic device and machine learning for automated recognition.
Main Results:
- Achieved attomolar detection limits with preamplification.
- Demonstrated a fast, highly sensitive, and single-pot multiplexing capability.
- Successfully applied CLAMP to detect human papillomavirus DNA in clinical samples.
Conclusions:
- CLAMP offers a robust platform for multiplexed CRISPR/Cas assays.
- The system streamlines detection through compartmentalization and automated analysis.
- CLAMP shows promise for clinical diagnostics, exemplified by HPV detection.
Keywords:
CRISPR/Cashuman papillomavirushydrogel microparticlesisothermal amplificationmultiplexed assays
