Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection
Abstract The CRISPR-based detection methods have been widely applied, yet they remain limited by the non-universal nature of one-pot diagnostic approaches. Here, we report a universal one-pot fluorescent method for the detection of epidemic pathogens, delivering results within 15-20 min. This method...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56516-3 |
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| author | Zhou-Hua Cheng Xi-Yan Luo Sheng-Song Yu Di Min Shu-Xia Zhang Xiao-Fan Li Jie-Jie Chen Dong-Feng Liu Han-Qing Yu |
| author_facet | Zhou-Hua Cheng Xi-Yan Luo Sheng-Song Yu Di Min Shu-Xia Zhang Xiao-Fan Li Jie-Jie Chen Dong-Feng Liu Han-Qing Yu |
| author_sort | Zhou-Hua Cheng |
| collection | DOAJ |
| description | Abstract The CRISPR-based detection methods have been widely applied, yet they remain limited by the non-universal nature of one-pot diagnostic approaches. Here, we report a universal one-pot fluorescent method for the detection of epidemic pathogens, delivering results within 15-20 min. This method uses heparin sodium to precisely tunes the cis-cleavage capability of Cas12 via interference with the Cas12a-crRNA binding process, thereby generating significant fluorescence due to the accumulation of isothermal amplification products. Additionally, this universal assay accommodates both classic and suboptimal PAMs, as well as various Cas12a subtypes such as LbCas12a, AsCas12a, and AapCas12b. Such a robust method demonstrates sensitivity and specificity exceeding 95% in the detection of monkeypox pseudovirus, influenza A virus, and SARS-CoV-2 from saliva or wastewater samples, when compared with qPCR or RT-qPCR. Moreover, the cost of heparin sodium per thousand uses is $0.01 to $0.04 only. Collectively, this universal and fast one-pot approach based on heparin sodium offers potential possibilities for point-of-care testing. |
| format | Article |
| id | doaj-art-179fb9edeaf74ccba8a5a312ace7f0ab |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-179fb9edeaf74ccba8a5a312ace7f0ab2025-02-02T12:32:37ZengNature PortfolioNature Communications2041-17232025-01-0116111610.1038/s41467-025-56516-3Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detectionZhou-Hua Cheng0Xi-Yan Luo1Sheng-Song Yu2Di Min3Shu-Xia Zhang4Xiao-Fan Li5Jie-Jie Chen6Dong-Feng Liu7Han-Qing Yu8CAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaFujian Institute of Hematology, Fujian Provincial Key Laboratory on Hematology, Fujian Medical University Union HospitalFujian Institute of Hematology, Fujian Provincial Key Laboratory on Hematology, Fujian Medical University Union HospitalCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaCAS Key Laboratory of Urban Pollutant Conversion, Department of Environmental Science and Engineering, University of Science and Technology of ChinaAbstract The CRISPR-based detection methods have been widely applied, yet they remain limited by the non-universal nature of one-pot diagnostic approaches. Here, we report a universal one-pot fluorescent method for the detection of epidemic pathogens, delivering results within 15-20 min. This method uses heparin sodium to precisely tunes the cis-cleavage capability of Cas12 via interference with the Cas12a-crRNA binding process, thereby generating significant fluorescence due to the accumulation of isothermal amplification products. Additionally, this universal assay accommodates both classic and suboptimal PAMs, as well as various Cas12a subtypes such as LbCas12a, AsCas12a, and AapCas12b. Such a robust method demonstrates sensitivity and specificity exceeding 95% in the detection of monkeypox pseudovirus, influenza A virus, and SARS-CoV-2 from saliva or wastewater samples, when compared with qPCR or RT-qPCR. Moreover, the cost of heparin sodium per thousand uses is $0.01 to $0.04 only. Collectively, this universal and fast one-pot approach based on heparin sodium offers potential possibilities for point-of-care testing.https://doi.org/10.1038/s41467-025-56516-3 |
| spellingShingle | Zhou-Hua Cheng Xi-Yan Luo Sheng-Song Yu Di Min Shu-Xia Zhang Xiao-Fan Li Jie-Jie Chen Dong-Feng Liu Han-Qing Yu Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection Nature Communications |
| title | Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection |
| title_full | Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection |
| title_fullStr | Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection |
| title_full_unstemmed | Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection |
| title_short | Tunable control of Cas12 activity promotes universal and fast one-pot nucleic acid detection |
| title_sort | tunable control of cas12 activity promotes universal and fast one pot nucleic acid detection |
| url | https://doi.org/10.1038/s41467-025-56516-3 |
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