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

• Published in: Nature communications 2024-10, Vol.15 (1), p.8560-16, Article 8560
• Main Authors: Guo, Yuqian, Zhou, Yaofeng, Duan, Hong, Xu, Derong, Wei, Min, Wu, Yuhao, Xiong, Ying, Chen, Xirui, Wang, Siyuan, Liu, Daofeng, Huang, Xiaolin, Xin, Hongbo, Xiong, Yonghua, Tang, Ben Zhong
• Format: Article
• Language: English
• Subjects: 639/301/1005/1009; 639/624/1107/510; 692/699/255/2514; 692/700/139/1512; Acids; Cleavage; COVID-19 - virology; CRISPR; CRISPR-Cas Systems; Diagnostic systems; DNA - genetics; Efficiency; Emission; Emissions; Fluorescence; Fluorescent Dyes - chemistry; Humanities and Social Sciences; Humans; Lighting; multidisciplinary; Norovirus - genetics; Nucleic acids; Portable equipment; SARS-CoV-2 - genetics; Science; Science (multidisciplinary); Severe acute respiratory syndrome coronavirus 2; Signal generation; Signal transduction; Single-stranded DNA
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-52931-0

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. Current CRISPR diagnostic approaches can be hampered by several limitations, including low signal transduction efficiency and limited sensitivity. Here, the authors present CrisprAIE, an approach based on CRISPR/Cas-mediated “one to more” lighting-up nucleic acid detection using aggregation-induced emission luminogens.