DNA test is widely used for disease detection and identification of GMOs.
However, DNA test is expensive and there is a need to develop a rapid,
low-cost, user-friendly, and field-deployable DNA test method. Huazhong
Agricultural University scientists developed a CRISPR Cas9-based DNA
detection method and the results are released in Molecular Breeding.
Fourteen (14) CRISPR RNAs were engineered to target two rice blast fungus
genes and a synthetic Cry1C gene used in Bt rice. With the use of a
fluorescent reporter, the targeted genes were easily detected LbCas12a after
recombinase polymerase amplification (RPA) for all CRISPR RNAs. Then the
researchers put together the filter paper-based DNA extraction and lateral
flow assay (LFA) with RPA-Cas12a for DNA detection. No extra instrumentation
was needed except for filter paper and LFA strip.
Rice blast pathogen and Bt rice were efficiently identified from leaf disc
samples using the modified DNA test method with highly active CRISPR RNAs.
Based on the findings, the optimized DNA test method could be applied in
disease diagnosis and GMO administration.