CRISPR-Cas12a-Based DNA Detection for Fast Pathogen Diagnosis and GMO Test in Plants

DNA test is widely used in plant pathogen diagnosis and genetically modified organism (GMO) administration. To date, a low-cost, user-friendly, and field-deployable DNA test method with high accuracy and sensitivity is still limited. Recently, the RNA programmable nuclease of CRISPR-Cas is engineered as a new nucleic acid detection platform, providing a novel and promising DNA test strategy for in-field crop disease diagnosis and GMO identification. In this study, we describe an all-paper-based DNA test method using CRISPR-Cas12a. This method combines filter-paper-based DNA purification, recombinase polymerase amplification, target gene detection with Cas12a, and lateral flow assay. Owing to its simplicity, efficiency, robustness, and low-cost, this all-paper-based Cas12a DNA test method could be easily applied in field for crop disease diagnosis and GMO test.