Advances in the application of CRISPR-Cas technology in rapid detection of pathogen nucleic acid

Xiaoping Li; Jiaye Zhong; Haoyu Li; Yinbiao Qiao; Xiaolei Mao; Huayan Fan; Yiwu Zhong; Saber Imani; Shusen Zheng; Jianhui Li

doi:10.3389/fmolb.2023.1260883

Advances in the application of CRISPR-Cas technology in rapid detection of pathogen nucleic acid

Front Mol Biosci. 2023 Sep 21:10:1260883. doi: 10.3389/fmolb.2023.1260883. eCollection 2023.

Authors

Xiaoping Li^#^{1

2}, Jiaye Zhong^#¹, Haoyu Li^{3

4}, Yinbiao Qiao^{5

3}, Xiaolei Mao¹, Huayan Fan¹, Yiwu Zhong⁶, Saber Imani¹, Shusen Zheng^{5

3

4}, Jianhui Li^{5

3

7

4

6}

Affiliations

¹ Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province, Shulan International Medical College, Zhejiang Shuren University, Hangzhou, China.
² Faculty of Medicine, Macau University of Science and Technology, Avenida Wai Long Taipa, Macau, China.
³ NHC Key Laboratory of Combined Multi-Organ Transplantation, Hangzhou, China.
⁴ Jinan Microecological Biomedicine Shandong Laboratory, Jinan, China.
⁵ Department of Hepatobiliary and Pancreatic Surgery, Department of Liver Transplantation, Shulan (Hangzhou) Hospital, Zhejiang Shuren University School of Medicine, Hangzhou, China.
⁶ Zhejiang Chinese Medical University, Hangzhou, China.
⁷ The Organ Repair and Regeneration Medicine Institute of Hangzhou, Hangzhou, China.

^# Contributed equally.

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) are widely used as gene editing tools in biology, microbiology, and other fields. CRISPR is composed of highly conserved repetitive sequences and spacer sequences in tandem. The spacer sequence has homology with foreign nucleic acids such as viruses and plasmids; Cas effector proteins have endonucleases, and become a hotspot in the field of molecular diagnosis because they recognize and cut specific DNA or RNA sequences. Researchers have developed many diagnostic platforms with high sensitivity, high specificity, and low cost by using Cas proteins (Cas9, Cas12, Cas13, Cas14, etc.) in combination with signal amplification and transformation technologies (fluorescence method, lateral flow technology, etc.), providing a new way for rapid detection of pathogen nucleic acid. This paper introduces the biological mechanism and classification of CRISPR-Cas technology, summarizes the existing rapid detection technology for pathogen nucleic acid based on the trans cleavage activity of Cas, describes its characteristics, functions, and application scenarios, and prospects the future application of this technology.

Keywords: CRISPR-Cas; Cas12; Cas13; Cas14; Cas9; pathogen nucleic acid; rapid detection.

Publication types

Review

Grants and funding

The authors declare financial support was received for the research, authorship, and/or publication of this article. This study was financially supported by 2023 National Student Innovation and Entrepreneurship Training Program Project (Item Number: 202311842021X), Major Science and Technology Projects of Hainan Province (ZDKJ2019009), the Public Projects of Zhejiang Province (LGF21H030006), a Research Project of Jinan Microecological Biomedicine Shandong Laboratory (JNL-2022002A and JNL-2022023C), Key Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang province (KFJJ2023005). The funder did not participate in the designing, performing or reporting in the current study.