CRISPR Electrochemical Biosensor for Rapid Biomarker Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current electrochemistry-based biosensing platforms face challenges in accuracy and cost-effectiveness for rapid detection of disease biomarkers, particularly in developing early-diagnostic point-of-care systems.
Innovation Solution
A CRISPR electrochemical biosensing system (E-CRISPR) that utilizes Cas12a-crRNA duplex to recognize and cleave target nucleic acid strands, employing a nonspecific ssDNA reporter with an electrochemical tag for signal transduction, enabling detection of various analytes like nucleic acids, peptides, and proteins through a portable and cost-efficient platform.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If electrochemistry-based biosensing platforms are used for rapid detection of disease biomarkers, then rapid signal readout and portability are achieved, but accuracy is compromised
Solution Approach 1:
The patent introduces an intermediary CRISPR-Cas12a system between the target biomarker and the electrochemical detection. The Cas12a enzyme acts as a mediator that specifically recognizes and cleaves target nucleic acids, then activates collateral cleavage of reporter molecules. This intermediary mechanism enhances detection accuracy through specific molecular recognition while preserving the rapid electrochemical readout capability.
Solution Approach 2:
The patent replaces complex mechanical or optical detection systems with an electrochemical transduction system. By using electrochemical tags (such as methylene blue) that generate electrical signals upon cleavage, the system achieves rapid signal readout without the complexity and cost of optical instruments, while maintaining high accuracy through the specific CRISPR recognition mechanism.
2Measurement precision
If complex sensing systems are developed to improve detection accuracy, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The CRISPR-Cas12a system performs self-service through its intrinsic abilities: the Cas12a enzyme automatically recognizes target sequences via crRNA guidance, unwinds DNA helices using its helicase activity, and activates collateral cleavage without external intervention. This self-service mechanism simplifies the overall sensing platform by eliminating the need for complex sample preparation or multiple processing steps.
Solution Approach 2:
The patent segments the detection function into distinct modular components: the Cas12a-crRNA complex for target recognition, the ssDNA reporter with electrochemical tag for signal generation, and the electrochemical sensor for readout. This segmentation allows each component to be optimized independently and simplifies the overall system design and fabrication.
3Measurement precision
If specific target recognition is implemented to improve accuracy, then measurement precision is improved, but the system becomes less adaptable to different analytes
Solution Approach 1:
The patent implements universality through the modular CRISPR system design. The Cas12a enzyme and electrochemical detection platform remain constant, while only the crRNA sequence needs to be changed to target different nucleic acid sequences. This allows a single sensing platform to detect multiple different analytes including various viruses, bacteria, and genetic mutations by simply reprogramming the guide RNA.
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 E-CRISPR system provides a robust, cost-effective, and accurate method for detecting biomolecules, offering a broad dynamic range and high sensitivity, with the ability to differentiate between target and non-target sequences, and can be used for both nucleic acid and protein detection in complex matrices.
Implementation Method 1
the electron transfer process between the gold electrode and the redox active species on the ssDNA can be electrochemically initiated and transduced
Implementation Method 2
the complementarity (between crRNA and target) dependent cleavage activity can further be activated
Implementation Method 3
a thiol moiety to tether on the sensor surface in order to acquire the signal electrically
Data Source
AI summary
A CRISPR electrochemical biosensing system (E-CRISPR) for detection of analytes includes a disposable, micro-fabricated three-electrode sensor that includes a working electrode, a counter electrode, a reference electrode, and a nonspecific ssDNA reporter with an electrochemical tag for signal transduction tethered to a surface of the working electrode; and a Cas12a-crRNA duplex that is designed to specifically recognize and cleave target nucleic acid strand based on the protospacer adjacent motif (PAM) sequence of the target and crRNA sequence, wherein the PAM recognition depends on specific 5′ TTTN nucleic acid sequence located at an opposite strand of a recognition strand, and wherein only upon the recognition of the PAM sequence by the Cas protein, the Cas protein, acting as a DNA helicase, unwinds the target DNA.