Development of a rapid antiviral screening assay based on eGFP reporter virus of Mayaro virus

Li, Xiaodan; Zhang, Hongqing; Zhang, Yanan; Li, Jiaqi; Wang, Zhen; Deng, Cheng-Lin; Jardim, Ana Carolina Gomes; Terzian, Ana Carolina Bernardes; Nogueira, Maurício Lacerda; Zhang, Bo

doi:10.1016/j.antiviral.2019.05.013

Cited by 18 publications

(16 citation statements)

References 42 publications

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“…Reporter-expressing viruses represent a powerful tool for both basic research and translational studies (16,17,(33)(34)(35)(36). Several research groups, including ours, have previously described recombinant viruses expressing reporter genes to easily study the biology of viral infections, to evaluate the efficacy of antivirals or NAbs, and for in vivo studies in validated animal models (37-51).…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Chiem

Vasquez

Park

et al. 2021

J Virol

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The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen responsible of coronavirus disease 2019 (COVID-19), has devastated public health services and economies worldwide. Despite global efforts to contain the COVID-19 pandemic, SARS-CoV-2 is now found in over 200 countries and has caused an upward death toll of over 1 million human lives as of November 2020. To date, only one Food and Drug Administration (FDA)-approved therapeutic drug (Remdesivir) and a monoclonal antibody, MAb (Bamlanivimab) are available for the treatment of SARS-CoV-2. As with other viruses, studying SARS-CoV-2 requires the use of secondary approaches to detect the presence of the virus in infected cells. To overcome this limitation, we have generated replication-competent recombinant (r)SARS-CoV-2 expressing fluorescent (Venus or mCherry) or bioluminescent (Nluc) reporter genes. Vero E6 cells infected with reporter-expressing rSARS-CoV-2 can be easily detected via fluorescence or luciferase expression and display a good correlation between reporter gene expression and viral replication. Moreover, rSARS-CoV-2 expressing reporter genes have comparable plaque sizes and growth kinetics to those of wild-type virus, rSARS-CoV-2/WT. We used these reporter-expressing rSARS-CoV-2 to demonstrate their feasibility to identify neutralizing antibodies (NAbs) or antiviral drugs. Our results demonstrate that reporter-expressing rSARS-CoV-2 represent an excellent option to identify therapeutics for the treatment of SARS-CoV-2, where reporter gene expression can be used as valid surrogates to track viral infection. Moreover, the ability to manipulate the viral genome opens the feasibility of generating viruses expressing foreign genes for their use as vaccines for the treatment of SARS-CoV-2 infection. IMPORTANCE Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen that causes coronavirus disease 2019 (COVID-19), has significantly impacted the human health and economic status worldwide. There is an urgent need to identify effective prophylactics and therapeutics for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. The use of fluorescent- or luciferase-expressing reporter expressing viruses has significantly advanced viral research. Here, we generated recombinant (r)SARS-CoV-2 expressing fluorescent (Venus and mCherry) or luciferase (Nluc) reporter genes and demonstrate that they represent an excellent option to track viral infections in vitro. Importantly, reporter-expressing rSARS-CoV-2 display similar growth kinetics and plaque phenotype that their wild-type counterpart (rSARS-CoV-2/WT), demonstrating their feasibility to identify drugs and/or neutralizing antibodies (NAbs) for the therapeutic treatment of SARS-CoV-2. Henceforth, these reporter-expressing rSARS-CoV-2 can be used to interrogate large libraries of compounds and/or monoclonal antibodies (MAb), in high-throughput screening settings, to identify those with therapeutic potential against SARS-CoV-2.

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Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Chiem

Vasquez

Park

et al. 2021

J Virol

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show abstract

“…Reporter-expressing viruses represent a powerful tool for both basic research and translational studies (14,15,(31)(32)(33)(34). Several research groups, including ours, have previously described recombinant viruses expressing reporter genes to easily study the biology of viral infections, to evaluate the efficacy of antivirals or NAbs, and for in vivo studies in validated animal models (35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48)(49).…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of recombinant SARS-CoV-2 expressing reporter genes

Chiem

Vasquez

Park

et al. 2020

Preprint

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The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen responsible of coronavirus disease 2019 (COVID-19), has devastated public health services and economies worldwide. Despite global efforts to contain the COVID-19 pandemic, SARS-CoV-2 is now found in over 200 countries and has caused an upward death toll of over 1 million human lives as of November 2020. To date, only one Food and Drug Administration (FDA)-approved therapeutic drug (Remdesivir) and a monoclonal antibody, MAb (Bamlanivimab), but no vaccines, are available for the treatment of SARS-CoV-2. As with other viruses, studying SARS-CoV-2 requires the use of secondary approaches to detect the presence of the virus in infected cells. To overcome this limitation, we have generated replication-competent recombinant (r)SARS-CoV-2 expressing fluorescent (Venus or mCherry) or bioluminescent (Nluc) reporter genes. Vero E6 cells infected with reporter-expressing rSARS-CoV-2 can be easily detected via fluorescence or luciferase expression and display a good correlation between reporter gene expression and viral replication. Moreover, rSARS-CoV-2 expressing reporter genes have comparable plaque sizes and growth kinetics to those of wild-type virus, rSARS-CoV-2/WT. We used these reporter-expressing rSARS-CoV-2 to demonstrate their feasibility to identify neutralizing antibodies (NAbs) or antiviral drugs. Our results demonstrate that reporter-expressing rSARS-CoV-2 represent an excellent option to identify therapeutics for the treatment of SARS-CoV-2, where reporter gene expression can be used as valid surrogates to track viral infection. Moreover, the ability to manipulate the viral genome opens the feasibility of generating viruses expressing foreign genes for their use as vaccines for the treatment of SARS-CoV-2 infection.ImportanceSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the pathogen that causes coronavirus disease 2019 (COVID-19), has significantly impacted the human health and economic status worldwide. There is an urgent need to identify effective prophylactics and therapeutics for the treatment of SARS-CoV-2 infection and associated COVID-19 disease. The use of fluorescent- or luciferase-expressing reporter expressing viruses has significantly advanced viral research. Here, we generated recombinant (r)SARS-CoV-2 expressing fluorescent (Venus and mCherry) or luciferase (Nluc) reporter genes and demonstrate that they represent an excellent option to track viral infections in vitro. Importantly, reporter-expressing rSARS-CoV-2 display similar growth kinetics and plaque phenotype that their wild-type counterpart (rSARS-CoV-2/WT), demonstrating their feasibility to identify drugs and/or neutralizing antibodies (NAbs) for the therapeutic treatment of SARS-CoV-2. Henceforth, these reporter-expressing rSARS-CoV-2 can be used to interrogate large libraries of compounds and/or monoclonal antibodies (MAb), in high-throughput screening settings, to identify those with therapeutic potential against SARS-CoV-2.

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“…Cell-based antiviral assays that rely on the infection of susceptible cells with a virus are commonly used in the initial stage of alphavirus drug discovery [14,76,77]. The antiviral efficacy can be expressed as a reduction in virus-induced cytopathic effect (CPE), which can be scored microscopically and/or quantified using colorimetric assays [78,79].…”

Section: Cell Culture Modelsmentioning

confidence: 99%

“…The antiviral efficacy can be expressed as a reduction in virus-induced cytopathic effect (CPE), which can be scored microscopically and/or quantified using colorimetric assays [78,79]. Alternatively, when the cell type is not susceptible to the virus-induced CPE, the antiviral efficacy can be expressed as a reduction in fluorescence or luminescence using specific reporter viruses [76].…”

Section: Cell Culture Modelsmentioning

confidence: 99%

“…Arthritogenic alphaviruses infect a wide range of cell types and tissues, including monocytes and/or macrophages, dendritic cells, synovial and dermal fibroblasts, endothelial cells and muscle cells [80]. However, other cell types are most commonly used for antiviral screening, including African green monkey kidney (Vero) cells, baby hamster kidney (BHK) cells, HeLa cells, human fetal lung fibroblast (MRC-5) cells and human embryonic kidney 293 (HEK-293T) cells [76,77,81]. Although these cells have little to no clinical relevance to alphavirus infections, they are very suitable for high-throughput screening campaigns.…”

Section: Cell Culture Modelsmentioning

confidence: 99%

See 1 more Smart Citation

Antiviral drug discovery against arthritogenic alphaviruses: Tools and molecular targets

Abdelnabi

Jacobs

Neyts

2020

Biochemical Pharmacology

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Alphaviruses are (mainly) arthropod-borne viruses that belong to the family of the Togaviridae. Based on the disease they cause, alphaviruses are divided into an arthritogenic and an encephalitic group. Arthritogenic alphaviruses such as the chikungunya virus (CHIKV), the Ross River virus (RRV) and the Mayaro virus (MAYV) have become a serious public health concern in recent years. Epidemics are associated with high morbidity and the infections cause in many patients debilitating joint pain that can persist for months to years. The recent (2013-2014) introduction of CHIKV in the Americas resulted in millions of infected persons. Massive outbreaks of CHIKV and other arthritogenic alphaviruses are likely to occur in the future.Despite the worldwide (re-)emergence of these viruses, there are no antivirals or vaccines available for the treatment or prevention of infections with alphaviruses. It is therefore of utmost importance to develop antiviral strategies against these viruses. We here review the possible molecular targets in the replication cycle of these viruses for the development of antivirals. In addition, we provide an overview of the currently available in vitro systems and mouse infection models that can be used to assess the potential antiviral effect against these viruses.

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Development of a rapid antiviral screening assay based on eGFP reporter virus of Mayaro virus

Cited by 18 publications

References 42 publications

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Generation and Characterization of Recombinant SARS-CoV-2 Expressing Reporter Genes

Generation and Characterization of recombinant SARS-CoV-2 expressing reporter genes

Antiviral drug discovery against arthritogenic alphaviruses: Tools and molecular targets

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