CRISPR Detection System for Rapid Pathogen Identification
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Solution Overview
Problem
Current nucleic acid-based detection methods for pathogens and other targets are often labor-intensive, time-consuming, and not suitable for resource-limited settings, requiring improvements for cost-effectiveness, speed, sensitivity, and accuracy, especially for multiplex testing and use in diverse operating conditions.
Innovation Solution
The use of target-specific guide nucleic acids (gNAs) mediated nuclease systems, such as CRISPR/Cas system proteins, for detecting and identifying nucleic acids, where labeled gNA-nucleic acid complexes bind to targets, allowing for sensitive and specific signal detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional pathogen detection methods (cell culture, PCR, enzyme immunoassay) are used, then detection accuracy can be achieved, but the methods are labor-intensive and time-consuming (taking several hours to days)
Solution Approach 1:
The patent replaces conventional mechanical and chemical detection systems (PCR, enzyme immunoassay) with a CRISPR-based biological detection system that uses guide RNA to direct Cas proteins to target nucleic acids, producing detectable signals through programmable molecular recognition rather than labor-intensive procedural steps
Solution Approach 2:
The CRISPR detection system performs self-guided target recognition and signal generation through the programmable guide RNA-Cas protein complex that autonomously binds to complementary target sequences and produces detectable signals without requiring extensive manual intervention or infrastructure
2Adaptability or versatility
If conventional detection methods are used, then detection capability is achieved, but they require centralized labs and are not suitable for resource-limited settings
Solution Approach 1:
The patent describes portable and disposable device configurations for CRISPR detection that can be used in resource-limited settings without requiring centralized laboratory infrastructure, making the detection system accessible in field conditions
Solution Approach 2:
The CRISPR detection platform provides universal applicability across diverse operating conditions and settings through its programmable nature, allowing the same core system to detect various targets (pathogens, SNPs, toxins) by simply changing the guide RNA sequence rather than requiring different infrastructure
3Adaptability or versatility
If multiplex testing capability is added to detection methods, then comprehensive pathogen identification is achieved, but the complexity and cost increase
Solution Approach 1:
The patent implements multiplex detection by dividing the detection process into separate reaction channels or compartments, each containing guide RNA-Cas protein complexes targeted to different pathogens, allowing simultaneous detection of multiple targets in a single sample through parallelized molecular recognition events
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables rapid, accurate, and cost-effective detection of multiple targets in various conditions, including resource-limited settings, with the potential for portable and disposable devices, enhancing pathogen detection and identification capabilities.
Implementation Method 1
target-specific guide nucleic acids (gNAs) mediated nuclease systems, such as guide RNA (gRNA) mediated CPJSPR/Cas system proteins
Implementation Method 2
the nucleic acid and the plurality of the gNA-nucleic acid-guided nuclease system protein complexes are labeled; in some embodiments, they are both labeled prior to the contacting step; and in some embodiments, they are both labeled after the contacting step. In some embodiments, the nucleic acid comprises a first label and the gNA-nucleic acid-guided nuclease system protein complexes comprise a second label, wherein the first and second label comprise a donor/acceptor pair for fluorescent resonance energy transfer (FRET)
Data Source
AI summary
Provided herein are methods and compositions for the detection and identification of targets in a sample, using target-specific guide nucleic acids and nucleic acid-guided nuclease system proteins.