Biomolecular Sensors Based on CRISPR/Cas Gene Editing

The CRISPR/Cas system is an adaptive immune system widely found in most bacteria. The CRISPR/Cas system detection technology has three stages of adaptation, expression, and interference, which are tailored to convert the sequence information of target nucleic acids into detectable signals such as fluorescence and colorimetric values to achieve specific target detection. The CRISPR/Cas biosensing system built by Lifeasible is a universal platform for nucleic acid detection, which can be used in plant pathogen detection, genotyping, gene mutation detection, and single nucleotide polymorphism identification. The system is characterized by high sensitivity, specificity, speed, and low cost and can achieve multiple detections of target nucleic acids.

Figure 1. Schematic mechanism of CRISPR/Cas detection system.

Fungal, bacterial, and viral plant diseases are serious threats to crop production and quality safety, e.g., Fusarium graminearum and Fusarium verticillioides are responsible for maize ear rot, stem rot, and wheat blast. Rice black-streaked dwarf virus (RBSDV), on the other hand, migrates and spreads between rice, wheat, and maize hosts via Delphacodes striatella, seriously threatening rice, wheat, and maize production. We have established a plant pathogen detection system based on the CRISPR/Cas system, which is fast, accurate, and sensitive. We can provide technical support for early warning and green prevention and control of diseases.

• For red blotch disease, we developed a detection method based on the colorimetric recognition technology of CRISPR/Cas12a and gold nano-plasma and combined it with PCR amplification technology. The detection limit of this method for acids is much higher than that of the traditional PCR method, and accurate detection and analysis results can be obtained even after multiple dilutions of nucleic acid samples.
• We have developed several sets of nucleic acid diagnostic techniques for plant pathogens based on the "trans-cutting" activity of CRISPR-Cas12a and CRISPR-Cas13 systems. Optimization of the sample processing method enables sensitive detection of dilutions of crude extracts of the samples. In addition, using Fusarium graminearum and Fusarium verticillioides complex infestation as samples, we have established a complex assay based on the combined application of Cas12/Cas13, which can diagnose multiple pathogens in a single assay and further expand the application scenarios and potentials of the assay tool.
• We detected polygalacturonase (PG) in dragon fruit Fusarium oxysporum using CRISPR/Cas9 technology.

Lifeasible's CRISPR/Cas-based biosensor system is based on the specific recognition of pathogen nucleic acid sequences by nucleic acids or probes. It amplifies biological signals and converts them into easily detectable physicochemical signals, which can be processed to achieve qualitative or quantitative detection of plant pathogens. Please feel free to contact us for more information about CRISPR/Cas system for plant pathogen detection.