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CRISPR Detection Service
Service Details
| Delivery Standards | Target plasmid RPA isothermal amplification primers Project report CRISPR reagent kit for 50 reactions |
|---|---|
| Applications | 1.Disease detection2.Food safety3.Animal disease diagnostics4.Environmental monitoring |
| Timeline/Pricing |
 Consult online for details |
FASST Rapid Detection Platform
FASST (Fast, Accurate, Specific, and Simple Test) is a next-generation, high-sensitivity, high-specificity, rapid isothermal single-tube nucleic acid detection technology based on CRISPR. Relying on the synergistic action of multiple enzymes at room temperature, FASST enables rapid nucleic acid amplification and detection. It offers dual specificity and dual signal amplification, making it a true point-of-care testing (POCT) technology.
Traditional CRISPR detection methods typically involve either two-tube or single-tube reactions. Two-tube reactions are complex and prone to contamination, while single-tube reactions, though simpler and with lower contamination risk, often suffer from low sensitivity and are time-consuming. The FASST technology, developed through EDITGENE's proprietary protein purification platform, overcomes these challenges by optimizing Cas enzyme structures and employing unique crRNA design logic. This approach enables the production of highly efficient crRNA and reporters, creating a single-tube detection system. It reduces contamination risk, improves detection sensitivity to the amol level, and shortens detection time to just 10 minutes, achieving highly sensitive, specific, rapid, and accurate nucleic acid detection, addressing the limitations of traditional CRISPR detection methods.
Schematic Diagram
Technical Advantages
Fast
Results in 5-20 minutes, achieving truly "rapid" detection
Sensitive
High-efficiency reaction, reaching detection limits at the amol level
Simple
Single-step sampling, easy to operate, constant temperature, and portable equipment.
Accurate
Dual specificity ensures precise target recognition
Technology Comparison
| Item | FASST | PCR | LAMP (Isothermal Amplification) | Traditional CRISPR Detection |
|---|---|---|---|---|
|
Reaction Temperature |
37-42℃ (Isothermal) |
95℃-55℃-72℃ (Variable) |
65℃ (Isothermal) |
37-42℃ (Isothermal) |
| Run Time | 5-20 mins | 1-2h | 40-60mins | 30-60 mins |
|
Primer Quantity |
2 primers |
2 primers |
4-6 primers |
2 primers |
| Reagent Form | Liquid/ Lyophilized | Liquid | Liquid | Liquid/Lyophilized |
| Equipment Requirement | Isothermal equipment (metal bath, water bath, etc.) | PCR Amplifier | Isothermal equipment | Isothermal equipment (metal bath, water bath, etc.) |
| Ease of Use | Extremely simple, truly portable | Requires professional operation, complex equipment | Simple to operate | Extremely simple, truly portable |
| Aerosol Contamination | Single-tube reaction, low contamination risk | Risk of aerosol contamination | Risk of aerosol contamination | Two-tube reaction, risk of aerosol contamination |
Services Provided by EDITGENE
EDITGENE offers the FASST rapid detection technology, a high-sensitivity, high-specificity, and fast isothermal nucleic acid detection platform developed based on CRISPR. FASST addresses the challenges of traditional CRISPR detection, such as complexity, contamination risks, low sensitivity, and long detection times. Leveraging our proprietary protein purification platform, we have optimized the Cas enzyme structure, developed a unique crRNA design logic, and produced highly efficient crRNAs. By using custom-designed reporters, we achieve higher sensitivity detection while simplifying operation and reducing contamination risks. Based on your experimental needs, you can choose between stepwise custom services or full-suite custom detection services.
Service Content and Delivery Standards
| Service Content | Service Description | Delivery Standard |
|---|---|---|
|
Preparation of Target Gene Template |
Synthesis of target plasmid | Target plasmid |
| Design and Synthesis of crRNA and RPA Isothermal Amplification Primers | Design of crRNA and RPA primers based on the target sequence | crRNA，2 OD |
| Activity Screening of RPA Isothermal Amplification Primers | Multiple RPA primers are used to amplify the target sequence, and the most efficient primer sequence is selected | RPA isothermal amplification primers, 2 OD |
| Activity Screening of crRNA | Multiple crRNAs are analyzed for activity, and the most efficient crRNA is identified | Final report and raw data |
| Establishment and Optimization of Experimental System for crRNA and RPA Isothermal Amplification Primers | Sensitivity, specificity, and accuracy tests for crRNA and RPA | Final report and raw data |
| FASST Full-suite Detection Services | / | Final report and CRISPR reagent kit for 50 reactions |
Service Workflow
Case Study
① African Swine Fever Virus Detection
Results: Using ASFV 1070, the FASST technology can detect African Swine Fever Virus, identifying 100-1000 copies of nucleic acid within 10 minutes, and 10-50 copies within 20 minutes.
② Parrot Bornavirus Detection
Results: The CRISPR/Cas12a two-tube detection method (30 minutes for RPA reaction and 10 minutes for CRISPR detection) can detect as few as 10 copies of Parrot Bornavirus nucleic acid within 40 minutes.
③ Brucella abortus Detection
Results: The CRISPR/Cas12a two-tube detection method (30 minutes for RPA reaction and 10 minutes for CRISPR detection) can detect as few as 100 copies of Brucella abortus nucleic acid within 40 minutes.
Advantage and Characteristic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
Optimazied Strategy
We have create a unique sgRNA Design Logic
FAQ
How to Improve the Detection Sensitivity of Cas Enzymes?
1.Design an efficient crRNA sequence. Proper design and structure prediction using online resources can help select suitable crRNA to achieve good trans-cleavage activity of the Cas enzyme.
2.Choose an appropriate signal reporter substrate. Research indicates that using a 15 nt single-stranded DNA (ssDNA) as a reporter substrate maximizes the cleavage reaction rate of Cas12a, significantly enhancing the reaction rate compared to the commonly used 5-nt ssDNA.
3.Optimize reaction conditions and buffers. Adjusting the CRISPR reaction parameters, such as the ratio of Cas enzyme to crRNA, the concentration of the Cas enzyme, and the reaction temperature, can improve detection performance to some extent.
2.Choose an appropriate signal reporter substrate. Research indicates that using a 15 nt single-stranded DNA (ssDNA) as a reporter substrate maximizes the cleavage reaction rate of Cas12a, significantly enhancing the reaction rate compared to the commonly used 5-nt ssDNA.
3.Optimize reaction conditions and buffers. Adjusting the CRISPR reaction parameters, such as the ratio of Cas enzyme to crRNA, the concentration of the Cas enzyme, and the reaction temperature, can improve detection performance to some extent.
How to Design crRNA?
1.The design process can follow these steps:
1.Identify the target gene sequence.
2.Specify the Cas protein being used. Different Cas proteins require corresponding PAM (Protospacer Adjacent Motif) sequences; for instance, Cas12a needs the "TTTV" PAM sequence for target recognition.
3.Select the crRNA targeting region. Choose a 20 nt nucleotide sequence on the target gene that is adjacent to the PAM site and pairs with the complementary strand of the crRNA.
4.Combine the selected 20 nt target sequence (variable part) with the scaffold sequence (fixed part) to design the crRNA sequence.
5.Use online tools such as CRISPR design tools (e.g., CRISPOR, Benchling, etc.) to assist in designing crRNA. These tools can predict the efficiency and specificity of the sgRNA, helping to avoid potential off-target effects.
6.After completing the design, the synthetic crRNA sequence can be ordered from a synthetic biology company.
1.Identify the target gene sequence.
2.Specify the Cas protein being used. Different Cas proteins require corresponding PAM (Protospacer Adjacent Motif) sequences; for instance, Cas12a needs the "TTTV" PAM sequence for target recognition.
3.Select the crRNA targeting region. Choose a 20 nt nucleotide sequence on the target gene that is adjacent to the PAM site and pairs with the complementary strand of the crRNA.
4.Combine the selected 20 nt target sequence (variable part) with the scaffold sequence (fixed part) to design the crRNA sequence.
5.Use online tools such as CRISPR design tools (e.g., CRISPOR, Benchling, etc.) to assist in designing crRNA. These tools can predict the efficiency and specificity of the sgRNA, helping to avoid potential off-target effects.
6.After completing the design, the synthetic crRNA sequence can be ordered from a synthetic biology company.
