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High-throughput multiplexed gene and cell doping analysis through CRISPR-Cas12a system integrated with blood direct
Joon-Yeop Yi1,2, Hyomin Choi3,4, Minyoung Kim1,2
1Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Korea.
Science Advances
|July 9, 2025
Summary
A new method called HiMDA uses CRISPR-Cas12a and direct PCR to detect gene and cell doping in blood. This high-throughput analysis is highly sensitive and can identify doping markers quickly.
Area of Science:
- Biotechnology
- Molecular Biology
- Forensic Science
Background:
- Gene and cell therapies present new doping challenges.
- Efficient and sensitive detection methods for gene and cell doping are needed.
Purpose of the Study:
- To develop a high-throughput multiplexed gene and cell doping analysis (HiMDA).
- To integrate CRISPR-Cas12a with blood direct polymerase chain reaction (PCR) for enhanced detection.
Main Methods:
- Utilized CRISPR-Cas12a system for sequence-specific DNA recognition.
- Employed blood direct PCR for simultaneous amplification of multiple exogenous genes from whole blood.
- Integrated a fluorescence reporter system for on-target detection.
Main Results:
- HiMDA requires only 5 microliters of blood and a 90-minute process.
- Achieved high sensitivity, detecting as few as 2.5 copies of doping target genes.
- Successfully identified in vivo doping up to 10 days post-administration.
- Demonstrated multiplexed, on-target detection of doping genes and cells.
Conclusions:
- HiMDA offers a robust, high-throughput solution for gene and cell doping detection.
- The method meets the sensitivity and selectivity demands for anti-doping research.
- HiMDA provides a flexible approach to address future doping detection challenges.
