Co-freezing localized CRISPR-Cas12a system enables rapid and sensitive nucleic acid analysis

Rapid and sensitive nucleic acid detection is vital in disease diagnosis and therapeutic assessment. Herein, we propose a co-freezing localized CRISPR-Cas12a (CL-Cas12a) strategy for sensitive nucleic acid detection. The CL-Cas12a was obtained through a 15-minute co-freezing process, allowing the Cas12a/crRNA complex and hairpin reporter confined on the AuNPs surface with high load efficiency, for rapid sensing of nucleic acid with superior performance to other localized Cas12a strategies. This CL-Cas12a based platform could quantitatively detect targets down to 98 aM in 30 min with excellent specificity. Furthermore, the CL-Cas12a successful applied to detect human papillomavirus infection and human lung cancer-associated single-nucleotide mutations. We also achieved powerful signal amplification for imaging Survivin mRNA in living cells. These findings highlight the potential of CL-Cas12a as an effective tool for nucleic acid diagnostics and disease monitoring.

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Publication Details

Journal
Journal of Nanobiotechnology
Published
2024-10-05
DOI
https://doi.org/10.1186/s12951-024-02831-8
Citations
13
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
3.04

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article

Co-freezing localized CRISPR-Cas12a system enables rapid and sensitive nucleic acid analysis

Siting Chen, Lei Zheng, Weilun Pan, Xiaohui Yan et al.
13 citations
Journal of Nanobiotechnology
CRISPR and Genetic Engineering
3.04
article

Co-freezing localized CRISPR-Cas12a system enables rapid and sensitive nucleic acid analysis

Siting Chen, Lei Zheng, Weilun Pan, Xiaohui Yan, Shihua Luo, Bo Situ, Chunchen Liu, Wenbin Li, Zuo‐Feng Zhang, Ling Li, Wanting Su, Zhang Ye
article en
13 citations

Abstract

Rapid and sensitive nucleic acid detection is vital in disease diagnosis and therapeutic assessment. Herein, we propose a co-freezing localized CRISPR-Cas12a (CL-Cas12a) strategy for sensitive nucleic acid detection. The CL-Cas12a was obtained through a 15-minute co-freezing process, allowing the Cas12a/crRNA complex and hairpin reporter confined on the AuNPs surface with high load efficiency, for rapid sensing of nucleic acid with superior performance to other localized Cas12a strategies. This CL-Cas12a based platform could quantitatively detect targets down to 98 aM in 30 min with excellent specificity. Furthermore, the CL-Cas12a successful applied to detect human papillomavirus infection and human lung cancer-associated single-nucleotide mutations. We also achieved powerful signal amplification for imaging Survivin mRNA in living cells. These findings highlight the potential of CL-Cas12a as an effective tool for nucleic acid diagnostics and disease monitoring.

Journal of NanobiotechnologyVol. 22(1)
Guangdong Medical College (CN), Nanfang Hospital (CN), Affiliated Hospital of Youjiang Medical University for Nationalities (CN), Southern Medical University (CN)
National Natural Science Foundation of China, Natural Science Foundation of Guangxi Province
Openalex Percentile: Top 8%
CRISPR and Genetic Engineering
3.04
DOIOpen Access PDFOpenAlex
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