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Review
. 2022;6(1):44-52.
doi: 10.1007/s41664-022-00212-4. Epub 2022 Feb 28.

Improved Strategies for CRISPR-Cas12-based Nucleic Acids Detection

Miao Qiu  1 , Xiao-Ming Zhou  2 , Lei Liu  1
Affiliations
Review

Improved Strategies for CRISPR-Cas12-based Nucleic Acids Detection

Miao Qiu et al. J Anal Test. 2022.

Abstract

The COVID-19 pandemic has brought great challenges to traditional nucleic acid detection technology. Thus, it is urgent to develop a more simple and efficient nucleic acid detection technology. CRISPR-Cas12 has signal amplification ability, high sensitivity and high nucleic acid recognition specificity, so it is considered as a nucleic acid detection tool with broad development prospects and high application value. This review paper discusses recent advances in CRISPR-Cas12-based nucleic acid detection, with an emphasis on the new research methods and means to improve the nucleic acid detection capability of CRISPR-Cas12. Strategies for improving sensitivity, optimization of integrated detection, development of simplified detection mode and improvement of quantitative detection capabilities are included. Finally, the future development of CRISPR-Cas12-based nucleic acids detection is prospected.

Keywords: CRISPR-Cas12; Integrated detection; Nucleic acid detection; Quantitative detection; Sensitivity; Simplified detection mode.

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Conflict of interest statement

Conflict of interestThe authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1
Integrated detection technology based on spatial–temporal separation. a Schematic of a CRISPR-Cas12-based nucleic acids detection technology integrating RPA with Cas12 detection in a single reaction system. Adapted from Ref. [27]. Copyright 2019 American Chemical Society. b Schematic of dynamic aqueous multiphase reaction (DAMR) system. Adapted from Ref. [33]. Copyright 2020 American Chemical Society
Fig. 2
Fig. 2
Nanoparticle-assisted CRISPR-Cas12-based nucleic acids detection. a Schematic of LFA which works in signal-off mode. Adapted from Ref. [48]. Copyright 2020 Elsevier. b Schematic of a magnetic pull-down-assisted colorimetric method. Adapted from Ref. [51]. Copyright 2021 American Chemical Society. c Schematic of colorimetric analysis using DNA-AuNP probes as trans-cleavage substrates. Adapted from ref. [53]. Copyright 2021 American Chemical Society. d Schematic of CRISPR-Cas12a coupled with PtNP for visual detection on a volumetric bar-chart chip. Adapted from Ref. [54]. Copyright 2019 American Chemical Society
Fig. 3
Fig. 3
Quantitative detection technologies based on isothermal amplification. a Schematic of digitization-enhanced CRISPR/Cas-assisted one-pot virus detection (deCOViD). Adapted from Ref. [55]. Copyright 2021 Wiley–VCH GmbH. b Schematic of digital warm-start CRISPR (dWS-CRISPR). Adapted from Ref. [38]. Copyright 2021 Elsevier
Fig. 4
Fig. 4
Schematic of amplification-free quantitative detection. Adapted from Ref. [15]. Copyright 2021 American Chemical Society
See this image and copyright information in PMC

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