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Luciferase-based SARS-CoV-2 Replicon System for Studying Viral Replication and Discontinuous Transcriptionстатья Тезисы
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или ScopusДата последнего поиска статьи во внешних источниках: 1 декабря 2021 г.
- Авторы: Galkin Simon, Anisenko Andrey, Shadrina Olga, Gottikh Marina
- Журнал: Protein Science
- Том: 30
- Номер: S1
- Год издания: 2021
- Издательство: Cold Spring Harbor Laboratory Press
- Местоположение издательства: United States
- Первая страница: 59
- Последняя страница: 60
- DOI: 10.1002/pro.4191
- Аннотация: Coronavirus disease 2019 (COVID-19) is induced by the SARS-CoV-2 novel betacoronavirus. Due to the COVID-19 pandemic, the WHO has declared a worldwide public health emergency. Almost 3 million covid-19 deaths were reported by the end of March 2021. Despite the recent advances in vaccine development, COVID-19 remains a serious threat due to controversial information about immunity duration and herd immunity attainability. SARS-CoV-2 is a (+) single-stranded RNA virus containing multiple open reading frames in its genome. During viral transcription, shorter subgenomic RNAs (sgRNAs) are produced. These RNAs are used to translate viral 3' distal genes. Our knowledge about the molecular basis of SARS-CoV-2 replication and discontinuous transcription is still limited. In particular, it is unknown whether or which cellular proteins are involved in these processes. Largely, it is due to the lack of simple and safe replicon systems, which allow the performing of highthroughput screenings. In this work, we have developed a reporter replicon system consisting of 5' coronaviral untranslated region (UTR), Firefly luciferase (Fluc), Puromycin resistance gene, Transcriptional RegulationSequence of SARS-CoV-2 essential for viral RNAdependent RNA polymerase jump, Renila luciferase (Rluc), and 3' coronaviral UTR. If all conditions for discontinuous transcription are met an increased Rluc/Fluc signal ratio is registered. We have identified a minimal list of viral proteins from orf1a/b sufficient for discontinuoustranscription. The created system can be used forthe high-throughput screening of cellular proteins and drugs affecting SARS-CoV-2 replication and sgRNAs synthesis. The work was supported by RFBR grant 20-04-60216.
- Добавил в систему: Готтих Марина Борисовна
Прикрепленные файлы
| № | Имя | Описание | Имя файла | Размер | Добавлен |
|---|---|---|---|---|---|
| 1. | Protein_Science.pdf | Protein_Science.pdf | 538,7 КБ | 24 ноября 2021 [gottikh] |