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...
Main Authors: | , , , , , , |
---|---|
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 |
id |
doaj-a577703c0599455ca3539c61760e5773 |
---|---|
record_format |
Article |
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 |
work_keys_str_mv |
AT qiangao acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhenwang acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhenlongliu acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT xiaoyuli acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT yongxinzhang acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhizhenzhang acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT shancen acellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT qiangao cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhenwang cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhenlongliu cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT xiaoyuli cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT yongxinzhang cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT zhizhenzhang cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus AT shancen cellbasedhighthroughputapproachtoidentifyinhibitorsofinfluenzaavirus |
_version_ |
1725771952842342400 |