CRISPR-enabled Multimodal Electrochemical Devices for Ultrasensitive, Preamplification-free Nucleic Acid Testing

This project aims to develop a next-generation, ultrasensitive, and easy-to-use diagnostic platform for detecting nucleic acids – biological molecules that serve as essential markers for diseases and pathogens. The proposed diagnostic device merges cutting-edge advances in nanomaterials, sensor engineering, and gene-editing-inspired detection to allow accurate and rapid identification of nucleic acids without the need for complex laboratory procedures. Unlike conventional approaches that require nucleic acid amplification and trained personnel, this platform enables direct detection at the point of care with performance rivaling gold-standard methods. The anticipated outcomes support the national interest by promoting scientific progress and public health preparedness, with potential applications in clinical diagnostics, environmental monitoring, food safety, and biodefense. This project will also contribute to workforce development through educational integration at multiple levels. Research outcomes will be incorporated into university-level engineering courses and used to develop hands-on learning modules for outreach programs. These activities aim to foster greater engagement in science and engineering fields through learning opportunities and exposure to interdisciplinary research. The goal of this project is to develop a preamplification-free electrochemical nucleic acid testing platform that combines graphene-based redox cycling with CRISPR-Cas enzymatic amplification to achieve attomolar-level sensitivity and high specificity. The research focuses on four core innovations: (1) signal amplification through redox-tagged reporter aptamers confined in a dual-electrode array of graphene microstructures; (2) modulation of redox cycling efficiency through plasma-induced functionalization of graphene to enhance charge transfer kinetics; (3) integration of 3D dual-functional capture electrodes that enable rapid and multimodal signal readout; and (4) demonstration of the platform for virus DNA detection using a CRISPR-Cas12a assay. While the initial focus is on detecting single-stranded DNA, the platform is adaptable to detect RNA targets through modification of the CRISPR assay. The project will advance the understanding of device-material-biomolecule interactions and provide a framework for the development of high-performance, field-deployable biosensors for a broad range of applications. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.