A cell-based, infectious-free, platform to identify inhibitors of lassa virus ribonucleoprotein (vRNP) activity
| Autor: | Catherine Z. Chen, Benson Yh. Cheng, Emilio Ortiz-Riaño, Juan Carlos de la Torre, Beatrice Cubitt, Charles D. Yeh, Wei Zheng, N.O.E. Southall, Luis Martinez-Sobrido, Yu-Jin Kim |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
viruses 030106 microbiology Drug Evaluation Preclinical Gene Expression Biology medicine.disease_cause Lymphocytic choriomeningitis Antiviral Agents Article Virus Small Molecule Libraries Viral Proteins 03 medical and health sciences chemistry.chemical_compound RNA interference Virology Chlorocebus aethiops medicine Animals Humans Lassa virus Lassa fever Vero Cells Ribonucleoprotein Pharmacology Dose-Response Relationship Drug Ribavirin High-Throughput Nucleotide Sequencing Reproducibility of Results medicine.disease 3. Good health HEK293 Cells 030104 developmental biology Ribonucleoproteins chemistry Vero cell RNA Interference Genetic Engineering |
| Zdroj: | Antiviral Research |
| ISSN: | 0166-3542 |
| DOI: | 10.1016/j.antiviral.2019.104667 |
| Popis: | The mammarenavirus Lassa (LASV) is highly prevalent in West Africa where it infects several hundred thousand individuals annually resulting in a high number of Lassa fever (LF) cases, a febrile disease associated with high morbidity and significant mortality. Mounting evidence indicates that the worldwide-distributed prototypic mammarenavirus lymphocytic choriomeningitis virus (LCMV) is a neglected human pathogen of clinical significance. There are not Food and Drug Administration (FDA) licensed vaccines and current anti-mammarenavirus therapy is limited to an off-label use of ribavirin that is only partially effective and can cause significant side effects. Therefore, there is an unmet need for novel antiviral drugs to combat LASV. This task would be facilitated by the implementation of high throughput screens (HTS) to identify inhibitors of the activity of the virus ribonucleoprotein (vRNP) responsible for directing virus RNA genome replication and gene transcription. The use of live LASV for this purpose is jeopardized by the requirement of biosafety level 4 (BSL4) containment. We have developed a virus-free cell platform, where expression levels of reporter genes serve as accurate surrogates of vRNP activity, to develop cell-based assays compatible with HTS to identify inhibitors of LASV and LCMV mammarenavirus vRNP activities. Highlights • Generation of cell lines harboring functional viral ribonucleoprotein complexes (vRNP) of LCMV and LASV. • LASV and LCMV vRNP-expressing cell-based screening assays for inhibitors of LASV and LCMV RNA replication and gene transcription. • Ionophore antibiotics narasin and valinomycin are potent inhibitors of LCMV and LASV vRNPs. • Inhibitors of mitochondrial electron transport complex (mETC) I and III are potent inhibitors of LCMV and LASV vRNPs. |
| Databáze: | OpenAIRE |
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