A cell-based high-throughput approach to identify inhibitors of influenza A virus

Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesize...

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Main Authors: Qian Gao, Zhen Wang, Zhenlong Liu, Xiaoyu Li, Yongxin Zhang, Zhizhen Zhang, Shan Cen
Format: Article
Language:English
Published: Elsevier 2014-08-01
Series:Acta Pharmaceutica Sinica B
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2211383514000574
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spelling doaj-a577703c0599455ca3539c61760e57732020-11-24T22:21:09ZengElsevierActa Pharmaceutica Sinica B2211-38352211-38432014-08-014430130610.1016/j.apsb.2014.06.005A cell-based high-throughput approach to identify inhibitors of influenza A virusQian Gao0Zhen Wang1Zhenlong Liu2Xiaoyu Li3Yongxin Zhang4Zhizhen Zhang5Shan Cen6Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaInstitute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaInstitute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaInstitute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaInstitute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaDepartment of Biochemistry, Guangdong Medical College, Zhanjiang, 524023, ChinaInstitute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaInfluenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA, which expresses Gaussia luciferase upon influenza A virus infection. Using this cell line, an assay was developed and optimized to search for inhibitors of influenza virus replication. Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format (Z′ factor value>0.8). A pilot screening provides further evidence for validation of the assay. Taken together, this work provides a simple, convenient, and reliable HTS assay to identify compounds with anti-influenza activity.http://www.sciencedirect.com/science/article/pii/S2211383514000574Cell-based assayGaussia luciferaseInfluenza A virusHigh-throughput screen
collection DOAJ
language English
format Article
sources DOAJ
author Qian Gao
Zhen Wang
Zhenlong Liu
Xiaoyu Li
Yongxin Zhang
Zhizhen Zhang
Shan Cen
spellingShingle Qian Gao
Zhen Wang
Zhenlong Liu
Xiaoyu Li
Yongxin Zhang
Zhizhen Zhang
Shan Cen
A cell-based high-throughput approach to identify inhibitors of influenza A virus
Acta Pharmaceutica Sinica B
Cell-based assay
Gaussia luciferase
Influenza A virus
High-throughput screen
author_facet Qian Gao
Zhen Wang
Zhenlong Liu
Xiaoyu Li
Yongxin Zhang
Zhizhen Zhang
Shan Cen
author_sort Qian Gao
title A cell-based high-throughput approach to identify inhibitors of influenza A virus
title_short A cell-based high-throughput approach to identify inhibitors of influenza A virus
title_full A cell-based high-throughput approach to identify inhibitors of influenza A virus
title_fullStr A cell-based high-throughput approach to identify inhibitors of influenza A virus
title_full_unstemmed A cell-based high-throughput approach to identify inhibitors of influenza A virus
title_sort cell-based high-throughput approach to identify inhibitors of influenza a virus
publisher Elsevier
series Acta Pharmaceutica Sinica B
issn 2211-3835
2211-3843
publishDate 2014-08-01
description Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA, which expresses Gaussia luciferase upon influenza A virus infection. Using this cell line, an assay was developed and optimized to search for inhibitors of influenza virus replication. Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format (Z′ factor value>0.8). A pilot screening provides further evidence for validation of the assay. Taken together, this work provides a simple, convenient, and reliable HTS assay to identify compounds with anti-influenza activity.
topic Cell-based assay
Gaussia luciferase
Influenza A virus
High-throughput screen
url http://www.sciencedirect.com/science/article/pii/S2211383514000574
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