Challenges and perspectives of CRISPR-based technology for diagnostic applications
Stefano Del Giovane, Neda Bagheri, Andrea C. Di Pede, Alejandro Chamorro-García, Simona Ranallo, Davide Migliorelli, Loïc Burr, Samantha Paoletti, Hatice Altug, Alessandro Porchetta
Abstract
The precision and versatility of CRISPR-based techniques, combined with the advantages of nucleic acid-based nanotechnology, hold great promise in transforming the landscape of molecular diagnostics. While significant progress has been made, current CRISPR-based platforms primarly focus on nucleic acid detection. To expand the applicability and fully leverage the advantages offered by CRISPR-based diagnostics, ongoing efforts explore molecular strategies to develop CRISPR sensors capable of detecting a diverse range of analytes beyond nucleic acids. In addition, challenges still persist in the adaptation of CRISPR platforms for point-of-care (POC) applications, involving concerns such as portability and automation, as well as the complexities associated with multiplexing. Here, we provide a detailed classification and comprehensive discussion of molecular strategies facilitating the conversion of non-nucleic acid target binding into CRISPR-powered outputs with an emphasis on their corresponding design principles. Furthermore, the second part of the review outlines current challenges and potential solutions for seamlessly integrating these strategies into user-friendly platforms and rapid tests specifically tailored for point-of-care (POC).