Clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostics have emerged as next-generation molecular diagnostics. In CRISPR-based diagnostics, Cas12 and Cas13 proteins have been widely employed to detect DNA and RNA, respectively. Herein, we developed a novel hybrid Cas protein capable of detecting universal nucleic acids (DNA and RNA). The CRISPR/hybrid Cas system simultaneously recognizes both DNA and RNA, enabling the dual detection of pathogenic viruses in a single tube. Using wild-type (WT) and N501Y mutant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as detection models, we successfully detected both virus strains with a detection limit of 10 viral copies per reaction without cross-reactivity. Furthermore, it is demonstrated the detection of WT SARS-CoV-2 and N501Y mutant variants in clinical samples by using the CRISPR/hybrid Cas system. The hybrid Cas protein is expected to be utilized in a molecular diagnostic method for infectious diseases, tissue and liquid biopsies, and other nucleic acid biomarkers.
Bibliographical noteFunding Information:
This research was supported by National R&D Programs through National Research Foundation (NRF) of Korea funded by Ministry of Science and ICT (MSIT) of Korea ( NRF-2021M3E5E3080379 , NRF-2021M3H4A1A02051048 , NRF-2018M3A9E2022821 , NRF-2021M3E5E3080844 , NRF-2022R1C1C1008815 , and NRF-2020R1A2C1010453 ), Korea Evaulation Institute of Industrial Technology (KEIT) grant funded by the Korea government ( MOTIE ) (No. RS-2022-00154853 ), Technology Development Program for Biological Hazards Management in Indoor Air through Korea Environment Industry & Technology Institute (KEITI) funded by Ministry of Environment (ME) of Korea ( 2021003370003 ), Nanomedical Devices Development Program of National Nano Fab Center ( CSM2105M101 ), and KRIBB Research Initiative Program ( 1711134081 ).
© 2022 The Authors
All Science Journal Classification (ASJC) codes
- Biomedical Engineering