CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection
Viruses, known for causing widespread biological harm and even extinction, pose significant challenges to public health. Virus detection is crucial for accurate disease diagnosis and preventing the spread of infections. Recently, the outstanding analytical performance of CRISPR/Cas biosensors has sh...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/29/24/6066 |
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| author | Zhenlin Shang Sitong Liu Dongxu Liu Xiaojing Pei Shujing Li Yifan He Yigang Tong Guoqi Liu |
| author_facet | Zhenlin Shang Sitong Liu Dongxu Liu Xiaojing Pei Shujing Li Yifan He Yigang Tong Guoqi Liu |
| author_sort | Zhenlin Shang |
| collection | DOAJ |
| description | Viruses, known for causing widespread biological harm and even extinction, pose significant challenges to public health. Virus detection is crucial for accurate disease diagnosis and preventing the spread of infections. Recently, the outstanding analytical performance of CRISPR/Cas biosensors has shown great potential and they have been considered as augmenting methods for reverse-transcription polymerase chain reaction (RT-PCR), which was the gold standard for nucleic acid detection. We herein utilized Cas12a with universal CRISPR RNA (crRNA) for indiscriminate virus detection by attaching the target to a longer track strand for isothermal amplification. The amplified products contain a domain that is recognized by the Cas12a/crRNA complex, triggering the cleavage of surrounding reporters to produce signals, thereby escaping the target dependence of crRNA recognition. The proposed method allows the same crRNA to detect multiple viral nucleic acids with high sensitivity, including but not limited to <i>SARS-CoV-2</i>, human papillomaviruses (<i>HPV</i>), <i>HCOV-NL63</i>, <i>HCOV-HKU1</i>, and miRNA biomarkers. Taking <i>SARS-CoV-2</i> and <i>HPV16</i> pseudoviruses as examples, this method was proved as a versatile and sensitive platform for molecular diagnostic applications. |
| format | Article |
| id | doaj-art-a37d9c301aeb4db79ed86e0db7b17b6a |
| institution | DOAJ |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-a37d9c301aeb4db79ed86e0db7b17b6a2025-08-20T02:43:53ZengMDPI AGMolecules1420-30492024-12-012924606610.3390/molecules29246066CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus DetectionZhenlin Shang0Sitong Liu1Dongxu Liu2Xiaojing Pei3Shujing Li4Yifan He5Yigang Tong6Guoqi Liu7School of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaSchool of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaSchool of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaSchool of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaSchool of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaSchool of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing 100048, ChinaCollege of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, ChinaBiotecnovo (Beijing) Co., Ltd., Room 801 Suit C Hengtai Center, Building 3 Gate, 18 North Feng Road, Fengtai District, Beijing 100176, ChinaViruses, known for causing widespread biological harm and even extinction, pose significant challenges to public health. Virus detection is crucial for accurate disease diagnosis and preventing the spread of infections. Recently, the outstanding analytical performance of CRISPR/Cas biosensors has shown great potential and they have been considered as augmenting methods for reverse-transcription polymerase chain reaction (RT-PCR), which was the gold standard for nucleic acid detection. We herein utilized Cas12a with universal CRISPR RNA (crRNA) for indiscriminate virus detection by attaching the target to a longer track strand for isothermal amplification. The amplified products contain a domain that is recognized by the Cas12a/crRNA complex, triggering the cleavage of surrounding reporters to produce signals, thereby escaping the target dependence of crRNA recognition. The proposed method allows the same crRNA to detect multiple viral nucleic acids with high sensitivity, including but not limited to <i>SARS-CoV-2</i>, human papillomaviruses (<i>HPV</i>), <i>HCOV-NL63</i>, <i>HCOV-HKU1</i>, and miRNA biomarkers. Taking <i>SARS-CoV-2</i> and <i>HPV16</i> pseudoviruses as examples, this method was proved as a versatile and sensitive platform for molecular diagnostic applications.https://www.mdpi.com/1420-3049/29/24/6066virusesCRISPR-Casuniversal crRNAnucleic acid detection |
| spellingShingle | Zhenlin Shang Sitong Liu Dongxu Liu Xiaojing Pei Shujing Li Yifan He Yigang Tong Guoqi Liu CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection Molecules viruses CRISPR-Cas universal crRNA nucleic acid detection |
| title | CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection |
| title_full | CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection |
| title_fullStr | CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection |
| title_full_unstemmed | CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection |
| title_short | CRISPR/Cas12a with Universal crRNA for Indiscriminate Virus Detection |
| title_sort | crispr cas12a with universal crrna for indiscriminate virus detection |
| topic | viruses CRISPR-Cas universal crRNA nucleic acid detection |
| url | https://www.mdpi.com/1420-3049/29/24/6066 |
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