4.6 Review

CRISPR-Cas13a system: A novel tool for molecular diagnostics

Journal

FRONTIERS IN MICROBIOLOGY
Volume 13, Issue -, Pages -

Publisher

FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2022.1060947

Keywords

CRISPR-Cas; CRISPR; Cas13a; biosensor; SHERLOCK; molecular diagnostics

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Funding

  1. National Natural Science Foundation of China
  2. Science Foundation of AMU
  3. [81570497]
  4. [21QNPY004]

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The CRISPR/Cas13a system, as a natural adaptive immune system, has been widely studied for its applications in molecular diagnostics, gene therapy, gene editing, and RNA imaging. It has great potential in the field of molecular diagnostics, with applications in pathogen detection, biomarker detection, and non-nucleic acid target detection.
The clustered regularly interspaced short palindromic repeats (CRISPR) system is a natural adaptive immune system of prokaryotes. The CRISPR-Cas system is currently divided into two classes and six types: types I, III, and IV in class 1 systems and types II, V, and VI in class 2 systems. Among the CRISPR-Cas type VI systems, the CRISPR/Cas13a system has been the most widely characterized for its application in molecular diagnostics, gene therapy, gene editing, and RNA imaging. Moreover, because of the trans-cleavage activity of Cas13a and the high specificity of its CRISPR RNA, the CRISPR/Cas13a system has enormous potential in the field of molecular diagnostics. Herein, we summarize the applications of the CRISPR/Cas13a system in the detection of pathogens, including viruses, bacteria, parasites, chlamydia, and fungus; biomarkers, such as microRNAs, lncRNAs, and circRNAs; and some non-nucleic acid targets, including proteins, ions, and methyl groups. Meanwhile, we highlight the working principles of some novel Cas13a-based detection methods, including the Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) and its improved versions, Cas13a-based nucleic acid amplification-free biosensors, and Cas13a-based biosensors for non-nucleic acid target detection. Finally, we focus on some issues that need to be solved and the development prospects of the CRISPR/Cas13a system.

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