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

被引：29

作者：

Guk, Kyeonghye ^{[1
]}

Yi, Soyeon ^{[1
]}

Kim, Hyeran ^{[1
]}

Bae, Yoonji ^{[1
]}

Yong, Dongeun ^{[2
,3
]}

Kim, Sunjoo ^{[4
]}

Lee, Kyu-Sun ^{[1
,6
]}

Lim, Eun-Kyung ^{[1
,5
,6
]}

Kang, Taejoon ^{[1
,6
,7
]}

Jung, Juyeon ^{[1
,5
,6
,7
]}

机构：

[1] Korea Res Inst Biosci & Biotechnol KRIBB, Bionanotechnol Res Ctr, 125 Gwahak ro, Daejeon 34141, South Korea

[2] Yonsei Univ, Dept Lab Med, 50 1 Yonsei ro, Seoul 03722, South Korea

[3] Yonsei Univ, Res Inst Bacterial Resistance, 50 1 Yonsei ro, Seoul 03722, South Korea

[4] Gyeongsang Natl Univ, Coll Med, Republ Korea Dept Lab Med, Jinju 52727, South Korea

[5] UST, KRIBB Sch Biotechnol, Dept Nanobiotechnol, 217 Gajeong ro, Daejeon 34113, South Korea

[6] Sungkyunkwan Univ, Sch Pharm, 2066 Seobu ro, Suwon 16419, South Korea

[7] KRIBB, Bionanotechnol Res Ctr, Daejeon 34141, South Korea

来源：

BIOSENSORS & BIOELECTRONICS | 2023年 / 219卷

基金：

新加坡国家研究基金会;

关键词：

CRISPR; Cas; DNA; RNA; Fusion protein; Severe acute respiratory syndrome coronavirus; 2; ASSAY; SARS-COV-2;

D O I：

10.1016/j.bios.2022.114819

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

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.

引用

页数：7