CRISPR Diagnostic System Attomolar Sensitivity Point-of-Care
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Solution Overview
Problem
Current nucleic acid detection methods face challenges in achieving high sensitivity and specificity while being cost-effective and portable, limiting their usability in point-of-care settings for various healthcare applications.
Innovation Solution
A CRISPR-based nucleic acid detection system utilizing a CRISPR system with an effector protein and guide RNAs designed to bind target molecules, combined with an RNA-based masking construct and optional nucleic acid amplification reagents, to amplify and detect target nucleic acids or peptides in a sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If qPCR approaches are used for nucleic acid detection, then sensitivity is improved, but cost and device complexity increase, limiting portability
Solution Approach 1:
The patent replaces complex mechanical/qPCR instrumentation with a CRISPR-based biochemical system that uses guide RNAs and effector proteins to detect nucleic acids through specific binding and cleavage mechanisms, enabling portability while maintaining sensitivity
Solution Approach 2:
The invention changes the detection mechanism from amplification-based (qPCR) to binding/cleavage-based (CRISPR), fundamentally altering how detection sensitivity is achieved without requiring complex thermal cycling instrumentation
2Ease of operation
If isothermal nucleic acid amplification with portable platforms is used, then portability is improved, but detection sensitivity decreases
Solution Approach 1:
The patent incorporates pre-amplified nucleic acid templates into the portable CRISPR detection device, allowing the system to achieve high sensitivity without requiring complex amplification instrumentation at the point of care
Solution Approach 2:
The invention uses guide RNAs as intermediaries that bridge the target nucleic acid and the effector protein, enabling highly specific detection with simple portable devices through programmed RNA-RNA or RNA-DNA binding
3Measurement precision
If high sensitivity and specificity detection is achieved, then diagnostic accuracy is improved, but cost increases
Solution Approach 1:
The patent uses inexpensive, easily synthesized guide RNAs and CRISPR effector proteins that can be produced at low cost, replacing expensive antibodies or complex assay reagents while maintaining high specificity
Solution Approach 2:
The invention creates a universal CRISPR detection platform where the same effector protein system can detect different targets by simply changing the guide RNA sequence, reducing development costs across multiple applications
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
The system enables sensitive and specific detection of nucleic acids and peptides at attomolar levels, differentiating targets from non-targets based on single base pair differences, and can be used in point-of-care applications due to its portability and cost-effectiveness.
Implementation Method 1
a CRISPR system comprising an effector protein and one or more guide RNAs designed to bind to corresponding target molecules
Implementation Method 2
activating the CRISPR effector protein via binding of the one or more guide RNAs to the one or more target molecules, wherein activating the CRISPR effector protein results in modification of the RNA-based masking construct such that a detectable positive signal is produced
Implementation Method 3
nucleic acid amplification reagents to amplify target RNA molecules in a sample
Data Source
AI summary
The embodiments disclosed herein utilized RNA targeting effectors to provide a robust CRISPR-based diagnostic with attomolar sensitivity. Embodiments disclosed herein can detect both DNA and RNA with comparable levels of sensitivity and can differentiate targets from non-targets based on single base pair differences. Moreover, the embodiments disclosed herein can be prepared in freeze-dried format for convenient distribution and point-of-care (POC) applications. Such embodiments are useful in multiple scenarios in human health including, for example, viral detection, bacterial strain typing, sensitive genotyping, and detection of disease-associated cell free DNA.