A CRISPR/Cas12a-Mediated Sensitive DNA Detection System for Gene-Edited Rice

Area of Science:

• Agricultural Biotechnology
• Molecular Biology
• Genetics

Background:

• Genome editing technologies like CRISPR/Cas9 enable diverse applications in transgenic research.
• Gene editing products may exhibit minimal genetic differences from conventional crops, complicating detection.
• Distinguishing gene-edited crops from conventional ones presents a significant analytical challenge.

Purpose of the Study:

• To establish a specific and sensitive CRISPR/Cas12a-mediated gene editing system for detecting target fragments in transgenic and commercial rice.
• To optimize a CRISPR/Cas12a visible detection system for accurate nucleic acid detection in gene-edited rice.
• To evaluate the specificity, sensitivity, and robustness of the CRISPR/Cas12a detection method.

Main Methods:

• Optimization of a CRISPR/Cas12a visible detection system for gene-edited rice.
• Utilization of gel electrophoresis and fluorescence-based methods for signal detection.
• Sitewise variant compact analysis to assess detection efficiencies of different base mutations.

Main Results:

• The developed CRISPR/Cas12a system demonstrated high precision, particularly for low-concentration samples.
• Single-base detection in gene-edited rice was achieved, with varying efficiencies for different mutations.
• The system successfully detected target fragments in both transgenic rice strains and commercial rice products, including those with multiple mutation types.

Conclusions:

• A robust CRISPR/Cas12a-mediated detection method for gene-edited rice has been developed.
• This method offers a new technical foundation for rapid field detection of gene-edited rice.
• The system provides specific, sensitive, and robust detection capabilities for gene-edited rice analysis.