CRISPR/Cas-mediated “one to more” lighting-up nucleic acid detection using aggregation-induced emission luminogens

Yuqian Guo, Yaofeng Zhou, Hong Duan, Derong Xu, Min Wei, Yuhao Wu, Ying Xiong, Xirui Chen, Siyuan Wang, Daofeng Liu, Xiaolin Huang (), Hongbo Xin, Yonghua Xiong () and Ben Zhong Tang ()
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Yuqian Guo: Nanchang University
Yaofeng Zhou: Nanchang University
Hong Duan: Beijing Technology & Business University
Derong Xu: Nanchang University
Min Wei: Nanchang University
Yuhao Wu: Nanchang University
Ying Xiong: Central South University of Forestry and Technology
Xirui Chen: Nanchang University
Siyuan Wang: China Agricultural University
Daofeng Liu: Jiangxi Provincial Center for Disease Control and Prevention
Xiaolin Huang: Nanchang University
Hongbo Xin: Nanchang University
Yonghua Xiong: Nanchang University
Ben Zhong Tang: The Chinese University of Hong Kong

Nature Communications, 2024, vol. 15, issue 1, 1-16

Abstract: Abstract CRISPR diagnostics are effective but suffer from low signal transduction efficiency, limited sensitivity, and poor stability due to their reliance on the trans-cleavage of single-stranded nucleic acid fluorescent reporters. Here, we present CrisprAIE, which integrates CRISPR/Cas reactions with “one to more” aggregation-induced emission luminogen (AIEgen) lighting-up fluorescence generated by the trans-cleavage of Cas proteins to AIEgen-incorporated double-stranded DNA labeled with single-stranded nucleic acid linkers and Black Hole Quencher groups at both ends (Q-dsDNA/AIEgens-Q). CrisprAIE demonstrates superior performance in the clinical nucleic acid detection of norovirus and SARS-CoV-2 regardless of amplification. Moreover, the diagnostic potential of CrisprAIE is further enhanced by integrating it with spherical nucleic acid-modified AIEgens (SNA/AIEgens) and a portable cellphone-based readout device. The improved CrisprAIE system, utilizing Q-dsDNA/AIEgen-Q and SNA/AIEgen reporters, exhibits approximately 80- and 270-fold improvements in sensitivity, respectively, compared to conventional CRISPR-based diagnostics. We believe CrisprAIE can be readily extended as a universal signal generation strategy to significantly enhance the detection efficiency of almost all existing CRISPR-based diagnostics.

Date: 2024
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DOI: 10.1038/s41467-024-52931-0

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