High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System
Programmed-1 ribosomal frameshifting (-1 PRF) is a translational mechanism adopted by some viruses, including SARS-CoV-2. To find a compound that can inhibit -1 PRF in SARS-CoV-2, we set up a high-throughput screening system using a HeLa cell extract-derived cell-free protein synthesis (CFPS) system...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-04-01
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| Series: | BioTechniques |
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| Online Access: | https://www.tandfonline.com/doi/10.2144/btn-2023-0102 |
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| author | Kodai Machida Rin Tanaka Seraya Miki Shotaro Noseda Mayumi Yuasa-Sunagawa Hiroaki Imataka |
| author_facet | Kodai Machida Rin Tanaka Seraya Miki Shotaro Noseda Mayumi Yuasa-Sunagawa Hiroaki Imataka |
| author_sort | Kodai Machida |
| collection | DOAJ |
| description | Programmed-1 ribosomal frameshifting (-1 PRF) is a translational mechanism adopted by some viruses, including SARS-CoV-2. To find a compound that can inhibit -1 PRF in SARS-CoV-2, we set up a high-throughput screening system using a HeLa cell extract-derived cell-free protein synthesis (CFPS) system. A total of 32,000 compounds were individually incubated with the CFPS system programmed with a -1 PRF-EGFP template. Several compounds were observed to decrease the -1 PRF-driven fluorescence, and one of them had some suppressive effect on -1 PRF of a SARS-CoV-2 genome sequence in transfected cells. Thus the CFPS system can be used as a tool for a high-throughput screening of chemicals. |
| format | Article |
| id | doaj-art-7aeabe3ef8ea485eb824b825ecaed003 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2024-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-7aeabe3ef8ea485eb824b825ecaed0032025-08-20T01:47:21ZengTaylor & Francis GroupBioTechniques0736-62051940-98182024-04-0176416116810.2144/btn-2023-0102High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis SystemKodai Machida0Rin Tanaka1Seraya Miki2Shotaro Noseda3Mayumi Yuasa-Sunagawa4Hiroaki Imataka5Department of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanDepartment of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanDepartment of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanDepartment of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanDepartment of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanDepartment of Applied Chemistry,Graduate School of Engineering, University of Hyogo, Himeji, 671-2201, JapanProgrammed-1 ribosomal frameshifting (-1 PRF) is a translational mechanism adopted by some viruses, including SARS-CoV-2. To find a compound that can inhibit -1 PRF in SARS-CoV-2, we set up a high-throughput screening system using a HeLa cell extract-derived cell-free protein synthesis (CFPS) system. A total of 32,000 compounds were individually incubated with the CFPS system programmed with a -1 PRF-EGFP template. Several compounds were observed to decrease the -1 PRF-driven fluorescence, and one of them had some suppressive effect on -1 PRF of a SARS-CoV-2 genome sequence in transfected cells. Thus the CFPS system can be used as a tool for a high-throughput screening of chemicals.https://www.tandfonline.com/doi/10.2144/btn-2023-0102cell-free protein synthesisdrug discoveryprogrammed-1 ribosomal frameshiftingSARS-CoV-2screening |
| spellingShingle | Kodai Machida Rin Tanaka Seraya Miki Shotaro Noseda Mayumi Yuasa-Sunagawa Hiroaki Imataka High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System BioTechniques cell-free protein synthesis drug discovery programmed-1 ribosomal frameshifting SARS-CoV-2 screening |
| title | High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System |
| title_full | High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System |
| title_fullStr | High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System |
| title_full_unstemmed | High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System |
| title_short | High-Throughput Screening for a SARS-CoV-2 Frameshifting Inhibitor Using a Cell-Free Protein Synthesis System |
| title_sort | high throughput screening for a sars cov 2 frameshifting inhibitor using a cell free protein synthesis system |
| topic | cell-free protein synthesis drug discovery programmed-1 ribosomal frameshifting SARS-CoV-2 screening |
| url | https://www.tandfonline.com/doi/10.2144/btn-2023-0102 |
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