{"title":"作为蛋白酶活性测量平台的水泡性口炎病毒","authors":"Stefanie Rauch, Francesco Costacurta, Dorothee von Laer, Emmanuel Heilmann","doi":"10.1002/cpz1.70062","DOIUrl":null,"url":null,"abstract":"<p>Protease inhibitors are among the most powerful antiviral drugs. They have been used successfully against viruses, such as the human immunodeficiency virus (HIV), hepatitis C virus (HCV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Protease inhibitor screening tools are therefore important to identify inhibitors that have the potential to become antiviral drugs. In this article, we describe newly developed cell- and virus replicon-based platforms to screen inhibitors. We developed the methods presented here by genetically modifying vesicular stomatitis virus, a model virus from the family <i>Rhabdoviridae</i>. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: M<sup>pro</sup>-On and -Off assay</p><p><b>Alternate Protocol 1</b>: Virus production with transient P- and L TransIT transfection</p><p><b>Alternate Protocol 2</b>: Virus production with transient P- and L Ca<sub>2</sub>PO<sub>4</sub> transfection</p><p><b>Alternate Protocol 3</b>: Luciferase-based variation of the On assay</p><p><b>Alternate Protocol 4</b>: Screening assay with fluorescence-activated cell sorting readout</p><p><b>Support Protocol</b>: Performing kinetic measurements with Off assay</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"4 11","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70062","citationCount":"0","resultStr":"{\"title\":\"Vesicular Stomatitis Virus as a Platform for Protease Activity Measurements\",\"authors\":\"Stefanie Rauch, Francesco Costacurta, Dorothee von Laer, Emmanuel Heilmann\",\"doi\":\"10.1002/cpz1.70062\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Protease inhibitors are among the most powerful antiviral drugs. They have been used successfully against viruses, such as the human immunodeficiency virus (HIV), hepatitis C virus (HCV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Protease inhibitor screening tools are therefore important to identify inhibitors that have the potential to become antiviral drugs. In this article, we describe newly developed cell- and virus replicon-based platforms to screen inhibitors. We developed the methods presented here by genetically modifying vesicular stomatitis virus, a model virus from the family <i>Rhabdoviridae</i>. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: M<sup>pro</sup>-On and -Off assay</p><p><b>Alternate Protocol 1</b>: Virus production with transient P- and L TransIT transfection</p><p><b>Alternate Protocol 2</b>: Virus production with transient P- and L Ca<sub>2</sub>PO<sub>4</sub> transfection</p><p><b>Alternate Protocol 3</b>: Luciferase-based variation of the On assay</p><p><b>Alternate Protocol 4</b>: Screening assay with fluorescence-activated cell sorting readout</p><p><b>Support Protocol</b>: Performing kinetic measurements with Off assay</p>\",\"PeriodicalId\":93970,\"journal\":{\"name\":\"Current protocols\",\"volume\":\"4 11\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-11-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70062\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70062\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70062","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Vesicular Stomatitis Virus as a Platform for Protease Activity Measurements
Protease inhibitors are among the most powerful antiviral drugs. They have been used successfully against viruses, such as the human immunodeficiency virus (HIV), hepatitis C virus (HCV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Protease inhibitor screening tools are therefore important to identify inhibitors that have the potential to become antiviral drugs. In this article, we describe newly developed cell- and virus replicon-based platforms to screen inhibitors. We developed the methods presented here by genetically modifying vesicular stomatitis virus, a model virus from the family Rhabdoviridae. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.
Basic Protocol: Mpro-On and -Off assay
Alternate Protocol 1: Virus production with transient P- and L TransIT transfection
Alternate Protocol 2: Virus production with transient P- and L Ca2PO4 transfection
Alternate Protocol 3: Luciferase-based variation of the On assay
Alternate Protocol 4: Screening assay with fluorescence-activated cell sorting readout
Support Protocol: Performing kinetic measurements with Off assay
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