CRISPR-electrochemical biosensors shift from signal-off to signal-on designs for improved sensitivity
This review evaluates the transition from signal-off to signal-on architectures in CRISPR-integrated electrochemical biosensors, noting that early turn-off designs are constrained by high background noise and limited sensitivity. Investigators outline how nanomaterials, DNA nanotechnology, and amplification strategies are being leveraged to build turn-on platforms that produce measurable electrochemical signals upon target recognition. The authors emphasize that while improving limits of detection and assay robustness remain critical engineering hurdles, these modular designs are well-positioned for decentralized and point-of-care molecular testing. The synthesis offers diagnostic developers a clear framework for optimizing signal transduction and advancing practical CRISPR-based electrochemical assays.
The original study
Incorporating CRISPR Techniques with Electrochemical Sensors: From Signal-Off to Signal-On Assays.
- Authors
- Yang C, Hu J, Zhang H, Le XC
- Journal
- ACS sensors
- Type
- Journal Article
- PMID
- 42696478
Original abstract
Electrochemical biosensors integrating the programmable nucleic acid recognition of CRISPR systems with the low-cost and portable electrochemical transduction have emerged as powerful sensing platforms for molecular diagnostics. Early designs mainly relied on turn-off signal transduction, where target-activated CRISPR enzymes cleave probes conjugated to the electrode, resulting in the release of redox reporters from the electrode surface and the consequent signal decrease. Although conceptually straightforward, these turn-off sensors are intrinsically limited by high background, low sensitivity, and large signal variations. To address these limitations, recent efforts have increasingly shifted toward turn-on strategies, in which electrochemical signals are generated in response to target binding. This review highlights recent advances in applying CRISPR technology to electrochemical biosensing, with a focus on the design principles of CRISPR systems and molecular assembly to achieve turn-off and turn-on signal transduction. Nanomaterials, DNA nanotechnology, and amplification strategies facilitate emerging turn-on approaches for sensitive electrochemical sensing. Key challenges and research needs include improving the limit of detection, robustness, and applicability to point-of-care and on-site testing. This review emphasizes the importance of signal-transduction designs and provides perspectives for developing sensitive, specific, and practical CRISPR-based electrochemical biosensors.