Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening
Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-12-01
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| Series: | SLAS Discovery |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472555224000510 |
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| author | Ryan Chan Christian Shema Mugisha Vorada Chuenchob Stephanie A. Moquin Ujjini H. Manjunatha Nadine Jarrousse Vineet D. Menachery Xuping Xie Erika L. Flannery Richard T. Eastman |
| author_facet | Ryan Chan Christian Shema Mugisha Vorada Chuenchob Stephanie A. Moquin Ujjini H. Manjunatha Nadine Jarrousse Vineet D. Menachery Xuping Xie Erika L. Flannery Richard T. Eastman |
| author_sort | Ryan Chan |
| collection | DOAJ |
| description | Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The identification of a novel alphacoronavirus canine CoV (CCoV-HuPn2018) in human patients in Malaysia underscores the potential for crossover infections to humans. The threat of the ever-evolving nature of viral infections as well as the lingering health and socioeconomic effects of the recent SARS-CoV-2 pandemic emphasize the urgent need for advanced antiviral drug screening tools that can be quickly implemented to strengthen preparedness and preventive measures against future outbreaks. Here, we present the development and validation of a novel RNA-fluorescence in situ hybridization (FISH) imaging assay as a 384-well, high-throughput rapid response platform for antiviral drug discovery. RNA-FISH is a powerful tool to visualize specific mRNA in cultured cells using a high-content imaging platform. The flexibility of RNA-FISH probe sets allows for the rapid design of viral genome-specific probes, enabling in vitro assay development to test for inhibition of viral replication by either biologic or small molecule inhibitors. Screening of 170 antiviral compounds in concentration-response demonstrates a strong correlation between the RNA-FISH assay and an immunofluorescence assay (IFA) for both human coronaviruses HCoV-OC43 and HCoV-229E. Additionally, we successfully applied this methodology in the context of CCoV strain 1–71, proving rapid development and deployment, opening new avenues for the evaluation of antiviral drugs to potential future emerging threats. |
| format | Article |
| id | doaj-art-d6e6474e81d84cbb9f2bab4c34ca05be |
| institution | Kabale University |
| issn | 2472-5552 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj-art-d6e6474e81d84cbb9f2bab4c34ca05be2024-12-17T05:00:16ZengElsevierSLAS Discovery2472-55522024-12-01298100189Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screeningRyan Chan0Christian Shema Mugisha1Vorada Chuenchob2Stephanie A. Moquin3Ujjini H. Manjunatha4Nadine Jarrousse5Vineet D. Menachery6Xuping Xie7Erika L. Flannery8Richard T. Eastman9Global Health, Biomedical Research, Novartis, Emeryville, CA 94608, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USADepartment of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston TX 77555, USA; Department of Pathology and Center for Biodefense & Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, TX 77555, USADepartment of Biochemistry & Molecular Biology, The University of Texas Medical Branch, Galveston, TX 77555, USAGlobal Health, Biomedical Research, Novartis, Emeryville, CA 94608, USA; Corresponding authors.Global Health, Biomedical Research, Novartis, Emeryville, CA 94608, USA; Corresponding authors.Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The identification of a novel alphacoronavirus canine CoV (CCoV-HuPn2018) in human patients in Malaysia underscores the potential for crossover infections to humans. The threat of the ever-evolving nature of viral infections as well as the lingering health and socioeconomic effects of the recent SARS-CoV-2 pandemic emphasize the urgent need for advanced antiviral drug screening tools that can be quickly implemented to strengthen preparedness and preventive measures against future outbreaks. Here, we present the development and validation of a novel RNA-fluorescence in situ hybridization (FISH) imaging assay as a 384-well, high-throughput rapid response platform for antiviral drug discovery. RNA-FISH is a powerful tool to visualize specific mRNA in cultured cells using a high-content imaging platform. The flexibility of RNA-FISH probe sets allows for the rapid design of viral genome-specific probes, enabling in vitro assay development to test for inhibition of viral replication by either biologic or small molecule inhibitors. Screening of 170 antiviral compounds in concentration-response demonstrates a strong correlation between the RNA-FISH assay and an immunofluorescence assay (IFA) for both human coronaviruses HCoV-OC43 and HCoV-229E. Additionally, we successfully applied this methodology in the context of CCoV strain 1–71, proving rapid development and deployment, opening new avenues for the evaluation of antiviral drugs to potential future emerging threats.http://www.sciencedirect.com/science/article/pii/S2472555224000510CoronavirusRNA-FISHHigh-content assayHigh-throughput screeningOC43229E |
| spellingShingle | Ryan Chan Christian Shema Mugisha Vorada Chuenchob Stephanie A. Moquin Ujjini H. Manjunatha Nadine Jarrousse Vineet D. Menachery Xuping Xie Erika L. Flannery Richard T. Eastman Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening SLAS Discovery Coronavirus RNA-FISH High-content assay High-throughput screening OC43 229E |
| title | Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening |
| title_full | Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening |
| title_fullStr | Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening |
| title_full_unstemmed | Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening |
| title_short | Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening |
| title_sort | rapid response rna fluorescence in situ hybridization fish assay platform for coronavirus antiviral high throughput screening |
| topic | Coronavirus RNA-FISH High-content assay High-throughput screening OC43 229E |
| url | http://www.sciencedirect.com/science/article/pii/S2472555224000510 |
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