Amplification-free GMO detection platform using crispr-based DNA sensor and microfluidic integration
A cost-effective CRISPR/Cas9 electrochemical sensor platform that detects plant gene mutations and GMO content without PCR amplification. Combines a stable glassy carbon electrode array in a microfluidic channel with single-base polymorphism sensitivity, achieving detection limits below 100 copies per microliter for point-of-collection plant genetics testing.
Overview
This solution offers an amplification-free DNA detection platform designed to identify genetically modified organisms (GMOs) and plant gene mutations with high sensitivity and specificity. By integrating CRISPR/Cas9 nucleic acid recognition with a microfluidic electrochemical sensor, the platform eliminates the need for PCR-based amplification, reducing cost, complexity, and turnaround time. The technology targets plant genetics research, agricultural biosecurity, and point-of-collection testing where rapid, equipment-light molecular diagnostics are needed.
Technical specifications
- CRISPR/Cas9 recognition: Uses programmable guide RNA (gRNA) to bind target DNA via Watson-Crick base pairing near a protospacer adjacent motif (PAM), enabling accurate discrimination of single-base polymorphisms with no cross-reactivity between highly homologous sequences.
- Electrochemical detection: Employs a stable, cost-effective glassy carbon electrode integrated into a microfluidic channel, with optional surface modification using materials such as graphene to further enhance sensitivity.
- Microelectrode array: Multiple CRISPR reagents targeting different genes can be immobilized on distinct electrodes within the array, supporting multiplexed detection in a single sample-to-answer workflow.
- Amplification-free performance: Demonstrated detection limit below 100 copies per microliter without nucleic acid amplification, validated against SARS-CoV-2 delta and omicron amplicons as a model for single-nucleotide specificity.
- Microfluidic integration: Sample preparation and processing occur within the microchannel, supporting automated handling and a streamlined workflow suitable for field or point-of-collection deployment.
Technology readiness level
The platform is at an early-to-mid stage of development, with core components individually validated. The CRISPR/Cas9 detection chemistry has demonstrated single-base specificity and sub-100 copies per microliter sensitivity without amplification. The glassy carbon electrode and microfluidic microsensor have been prototyped through collaboration between the Department of Mechanical Engineering and the Medical School at the University of Minnesota. A planned two-phase validation will integrate optimized CRISPR reagents with the sensor platform, calibrate the multiplexed electrode array, and conduct on-site testing with provided plant samples to assess real-world performance.
About University of Minnesota
The University of Minnesota is a flagship, comprehensive public research university spanning multiple campuses, with a large research enterprise and clinical integration. Industry engages through co-located labs on the Twin Cities campuses, access to an academic health system for clinical translation, and pilot and field-testing facilities that speed scale-up. A statewide extension network and outreach centers provide real-world sites and data partnerships across Minnesota, while proximity to a dense medtech and Fortune 500 corridor enables frequent collaboration. Research is supported by competitive federal funding, including NIH, NSF, DOE, USDA, and DoD. A dedicated technology transfer office manages IP, licensing, sponsored research agreements, and startup incubation to speed commercialization.
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