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Hybrid CRISPR/Cas protein for one-pot detection of DNA and RNA

Authors
 Kyeonghye Guk  ;  Soyeon Yi  ;  Hyeran Kim  ;  Yoonji Bae  ;  Dongeun Yong  ;  Sunjoo Kim  ;  Kyu-Sun Lee  ;  Eun-Kyung Lim  ;  Taejoon Kang  ;  Juyeon Jung 
Citation
 BIOSENSORS & BIOELECTRONICS, Vol.219 : 114819, 2023-01 
Journal Title
BIOSENSORS & BIOELECTRONICS
ISSN
 0956-5663 
Issue Date
2023-01
Keywords
CRISPR/Cas ; DNA ; Fusion protein ; RNA ; Severe acute respiratory syndrome coronavirus 2
Abstract
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 suc-cessfully 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.
Files in This Item:
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DOI
10.1016/j.bios.2022.114819
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Yong, Dong Eun(용동은) ORCID logo https://orcid.org/0000-0002-1225-8477
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/199687
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