Replication-Competent Reporter-Expressing Viruses

A special issue of Viruses (ISSN 1999-4915).

Deadline for manuscript submissions: closed (29 February 2016) | Viewed by 127087

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Special Issue Editor

1. Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14625, USA
2. Texas Biomedical Research Institute, San Antonio, TX 78245, USA
Interests: virology; vaccines; antivirals; influenza viruses; arenaviruses; Zika virus; coronavirus; SARS-CoV-2; COVID-19; innate immunity; adaptive immunity; interferon; virus-host interactions
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Special Issue Information

Dear Colleagues,

With the development of reverse genetics systems, recombinant viruses expressing reporter fluorescent or bioluminescent genes represent an excellent option to evaluate the dynamics of viral infection progression in both cultured cells and/or validated animal models of infection. Expression of reporter proteins allows for direct viral detection in vitro and in vivo, without the use of secondary methodologies to identify infected cells. By eliminating the need of secondary labeling, fluorescent or bioluminescence tractable replicating-compatible viruses provide an ideal tool to monitor viral infections in real time, representing a significant advance in the study of the biology of viruses, to evaluate vaccination approaches, and to identify new therapeutics against viral infections using high-through put screening settings. In this Special Issue, we aim to review replicating-competent, reporter-expressing viruses belonging to different families, methods of characterization, and applications to facilitate the study of in vitro and in vivo viral infections. Contrasting advantages, we also seek to discuss disadvantages associated with these reporter-expressing viruses. Finally, we will provide rational future perspectives and additional avenues for the development, characterization, and application of recombinant, reporter-expressing, competent viruses.

Prof. Dr. Luis Martinez-Sobrido 
Guest Editor

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Keywords

  • reporter genes
  • recombinant viruses
  • replicating-competent viruses
  • antivirals
  • vaccines
  • in vivo imaging
  • dynamics of viral infections

Published Papers (15 papers)

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Research

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20768 KiB  
Article
Development of Neutralization Assay Using an eGFP Chikungunya Virus
by Cheng-Lin Deng, Si-Qing Liu, Dong-Gen Zhou, Lin-Lin Xu, Xiao-Dan Li, Pan-Tao Zhang, Peng-Hui Li, Han-Qing Ye, Hong-Ping Wei, Zhi-Ming Yuan, Cheng-Feng Qin and Bo Zhang
Viruses 2016, 8(7), 181; https://doi.org/10.3390/v8070181 - 28 Jun 2016
Cited by 19 | Viewed by 7750
Abstract
Chikungunya virus (CHIKV), a member of the Alphavirus genus, is an important human emerging/re-emerging pathogen. Currently, there are no effective antiviral drugs or vaccines against CHIKV infection. Herein, we construct an infectious clone of CHIKV and an eGFP reporter CHIKV (eGFP-CHIKV) with an [...] Read more.
Chikungunya virus (CHIKV), a member of the Alphavirus genus, is an important human emerging/re-emerging pathogen. Currently, there are no effective antiviral drugs or vaccines against CHIKV infection. Herein, we construct an infectious clone of CHIKV and an eGFP reporter CHIKV (eGFP-CHIKV) with an isolated strain (assigned to Asian lineage) from CHIKV-infected patients. The eGFP-CHIKV reporter virus allows for direct visualization of viral replication through the levels of eGFP expression. Using a known CHIKV inhibitor, ribavirin, we confirmed that the eGFP-CHIKV reporter virus could be used to identify inhibitors against CHIKV. Importantly, we developed a novel and reliable eGFP-CHIKV reporter virus-based neutralization assay that could be used for rapid screening neutralizing antibodies against CHIKV. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Article
Development of a Triple-Color Pseudovirion-Based Assay to Detect Neutralizing Antibodies against Human Papillomavirus
by Jianhui Nie, Yangyang Liu, Weijin Huang and Youchun Wang
Viruses 2016, 8(4), 107; https://doi.org/10.3390/v8040107 - 25 Apr 2016
Cited by 28 | Viewed by 6131
Abstract
Pseudovirion-based neutralization assay is considered the gold standard method for evaluating the immune response to human papillomavirus (HPV) vaccines. In this study, we developed a multicolor neutralization assay to simultaneously detect the neutralizing antibodies against different HPV types. FluoroSpot was used to interpret [...] Read more.
Pseudovirion-based neutralization assay is considered the gold standard method for evaluating the immune response to human papillomavirus (HPV) vaccines. In this study, we developed a multicolor neutralization assay to simultaneously detect the neutralizing antibodies against different HPV types. FluoroSpot was used to interpret the fluorescent protein expression instead of flow cytometry. The results of FluoroSpot and flow cytometry showed good consistency, with R2 > 0.98 for the log-transformed IC50 values. Regardless of the reporter color, the single-, dual-, and triple-color neutralization assays reported identical results for the same samples. In low-titer samples from naturally HPV-infected individuals, there was strong agreement between the single- and triple-color assays, with kappa scores of 0.92, 0.89, and 0.96 for HPV16, HPV18, and HPV58, respectively. Good reproducibility was observed for the triple-color assay, with coefficients of variation of 2.0%–41.5% within the assays and 8.3%–36.2% between the assays. Three triple-color systems, HPV16-18-58, HPV6-33-45, and HPV11-31-52, were developed that could evaluate the immunogenicity of a nonavalent vaccine in three rounds of the assay. With the advantages of an easy-to-use procedure and less sample consumption, the multiple-color assay is more suitable than classical assays for large sero-epidemiological studies and clinical trials and is more amenable to automation. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Article
Recombinant Pseudorabies Virus (PRV) Expressing Firefly Luciferase Effectively Screened for CRISPR/Cas9 Single Guide RNAs and Antiviral Compounds
by Yan-Dong Tang, Ji-Ting Liu, Qiong-Qiong Fang, Tong-Yun Wang, Ming-Xia Sun, Tong-Qing An, Zhi-Jun Tian and Xue-Hui Cai
Viruses 2016, 8(4), 90; https://doi.org/10.3390/v8040090 - 29 Mar 2016
Cited by 28 | Viewed by 7421
Abstract
A Pseudorabies virus (PRV) variant has emerged in China since 2011 that is not protected by commercial vaccines, and has not been well studied. The PRV genome is large and difficult to manipulate, but it is feasible to use clustered, regularly interspaced short [...] Read more.
A Pseudorabies virus (PRV) variant has emerged in China since 2011 that is not protected by commercial vaccines, and has not been well studied. The PRV genome is large and difficult to manipulate, but it is feasible to use clustered, regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology. However, identification of single guide RNA (sgRNA) through screening is critical to the CRISPR/Cas9 system, and is traditionally time and labor intensive, and not suitable for rapid and high throughput screening of effective PRV sgRNAs. In this study, we developed a recombinant PRV strain expressing firefly luciferase and enhanced green fluorescent protein (EGFP) as a reporter virus for PRV-specific sgRNA screens and rapid evaluation of antiviral compounds. Luciferase activity was apparent as soon as 4 h after infection and was stably expressed through 10 passages. In a proof of the principle screen, we were able to identify several PRV specific sgRNAs and confirmed that they inhibited PRV replication using traditional methods. Using the reporter virus, we also identified PRV variants lacking US3, US2, and US9 gene function, and showed anti-PRV activity for chloroquine. Our results suggest that the reporter PRV strain will be a useful tool for basic virology studies, and for developing PRV control and prevention measures. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Article
Efficient Co-Replication of Defective Novirhabdovirus
by Ronan N. Rouxel, Emilie Mérour, Stéphane Biacchesi and Michel Brémont
Viruses 2016, 8(3), 69; https://doi.org/10.3390/v8030069 - 04 Mar 2016
Cited by 15 | Viewed by 5489
Abstract
We have generated defective Viral Hemorrhagic Septicemia Viruses (VHSV) which express either the green fluorescent protein (GFP) or a far-red fluorescent protein (mKate) by replacing the genes encoding the nucleoprotein N or the polymerase-associated P protein. To recover viable defective viruses, rVHSV-ΔN-Red and [...] Read more.
We have generated defective Viral Hemorrhagic Septicemia Viruses (VHSV) which express either the green fluorescent protein (GFP) or a far-red fluorescent protein (mKate) by replacing the genes encoding the nucleoprotein N or the polymerase-associated P protein. To recover viable defective viruses, rVHSV-ΔN-Red and rVHSV-ΔP-Green, fish cells were co-transfected with both deleted cDNA VHSV genomes, together with plasmids expressing N, P and L of the RNA-dependent RNA polymerase. After one passage of the transfected cell supernatant, red and green cell foci were observed. Viral titer reached 107 PFU/mL after three passages. Infected cells were always red and green with the very rare event of single red or green cell foci appearing. To clarify our understanding of how such defective viruses could be so efficiently propagated, we investigated whether (i) a recombination event between both defective genomes had occurred, (ii) whether both genomes were co-encapsidated in a single viral particle, and (iii) whether both defective viruses were always replicated together through a complementation phenomenon or even as conglomerate. To address these hypotheses, genome and viral particles have been fully characterized and, thus, allowing us to conclude that rVHSV-ΔN-Red and rVHSV-ΔP-Green are independent viral particles which could propagate only by simultaneously infecting the same cells. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Article
Fluorescent Protein-Tagged Sindbis Virus E2 Glycoprotein Allows Single Particle Analysis of Virus Budding from Live Cells
by Joyce Jose, Jinghua Tang, Aaron B. Taylor, Timothy S. Baker and Richard J. Kuhn
Viruses 2015, 7(12), 6182-6199; https://doi.org/10.3390/v7122926 - 27 Nov 2015
Cited by 23 | Viewed by 9382
Abstract
Sindbis virus (SINV) is an enveloped, mosquito-borne alphavirus. Here we generated and characterized a fluorescent protein-tagged (FP-tagged) SINV and found that the presence of the FP-tag (mCherry) affected glycoprotein transport to the plasma membrane whereas the specific infectivity of the virus was not [...] Read more.
Sindbis virus (SINV) is an enveloped, mosquito-borne alphavirus. Here we generated and characterized a fluorescent protein-tagged (FP-tagged) SINV and found that the presence of the FP-tag (mCherry) affected glycoprotein transport to the plasma membrane whereas the specific infectivity of the virus was not affected. We examined the virions by transmission electron cryo-microscopy and determined the arrangement of the FP-tag on the surface of the virion. The fluorescent proteins are arranged icosahedrally on the virus surface in a stable manner that did not adversely affect receptor binding or fusion functions of E2 and E1, respectively. The delay in surface expression of the viral glycoproteins, as demonstrated by flow cytometry analysis, contributed to a 10-fold reduction in mCherry-E2 virus titer. There is a 1:1 ratio of mCherry to E2 incorporated into the virion, which leads to a strong fluorescence signal and thus facilitates single-particle tracking experiments. We used the FP-tagged virus for high-resolution live-cell imaging to study the spatial and temporal aspects of alphavirus assembly and budding from mammalian cells. These processes were further analyzed by thin section microscopy. The results demonstrate that SINV buds from the plasma membrane of infected cells and is dispersed into the surrounding media or spread to neighboring cells facilitated by its close association with filopodial extensions. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Review

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2395 KiB  
Review
Fluorescent and Bioluminescent Reporter Myxoviruses
by Christina A. Rostad, Michael C. Currier and Martin L. Moore
Viruses 2016, 8(8), 214; https://doi.org/10.3390/v8080214 - 04 Aug 2016
Cited by 6 | Viewed by 4730
Abstract
The advent of virus reverse genetics has enabled the incorporation of genetically encoded reporter proteins into replication-competent viruses. These reporters include fluorescent proteins which have intrinsic chromophores that absorb light and re-emit it at lower wavelengths, and bioluminescent proteins which are luciferase enzymes [...] Read more.
The advent of virus reverse genetics has enabled the incorporation of genetically encoded reporter proteins into replication-competent viruses. These reporters include fluorescent proteins which have intrinsic chromophores that absorb light and re-emit it at lower wavelengths, and bioluminescent proteins which are luciferase enzymes that react with substrates to produce visible light. The incorporation of these reporters into replication-competent viruses has revolutionized our understanding of molecular virology and aspects of viral tropism and transmission. Reporter viruses have also enabled the development of high-throughput assays to screen antiviral compounds and antibodies and to perform neutralization assays. However, there remain technical challenges with the design of replication-competent reporter viruses, and each reporter has unique advantages and disadvantages for specific applications. This review describes currently available reporters, design strategies for incorporating reporters into replication-competent paramyxoviruses and orthomyxoviruses, and the variety of applications for which these tools can be utilized both in vitro and in vivo. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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1573 KiB  
Review
Reporter-Expressing, Replicating-Competent Recombinant Arenaviruses
by Luis Martínez-Sobrido and Juan Carlos De la Torre
Viruses 2016, 8(7), 197; https://doi.org/10.3390/v8070197 - 20 Jul 2016
Cited by 12 | Viewed by 8526
Abstract
Several arenaviruses cause hemorrhagic fever (HF) disease in humans and pose an important public health problem in their endemic regions. To date, no Food and Drug Administration (FDA)-licensed vaccines are available to combat human arenavirus infections, and current anti-arenaviral drug therapy is limited [...] Read more.
Several arenaviruses cause hemorrhagic fever (HF) disease in humans and pose an important public health problem in their endemic regions. To date, no Food and Drug Administration (FDA)-licensed vaccines are available to combat human arenavirus infections, and current anti-arenaviral drug therapy is limited to an off-label use of ribavirin that is only partially effective. The development of arenavirus reverse genetic approaches has provided investigators with a novel and powerful approach for the study of arenavirus biology including virus–host interactions underlying arenavirus induced disease. The use of cell-based minigenome systems has allowed examining the cis- and trans-acting factors involved in arenavirus replication and transcription, as well as particle assembly and budding. Likewise, it is now feasible to rescue infectious arenaviruses containing predetermined mutations in their genomes to investigate virus-host interactions and mechanisms of pathogenesis. The use of reverse genetics approaches has also allowed the generation of recombinant arenaviruses expressing additional genes of interest. These advances in arenavirus molecular genetics have also facilitated the implementation of novel screens to identify anti-arenaviral drugs, and the development of novel strategies for the generation of arenavirus live-attenuated vaccines. In this review, we will summarize the current knowledge on reporter-expressing, replicating-competent arenaviruses harboring reporter genes in different locations of the viral genome and their use for studying and understanding arenavirus biology and the identification of anti-arenaviral drugs to combat these important human pathogens. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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724 KiB  
Review
Recombinant Ranaviruses for Studying Evolution of Host–Pathogen Interactions in Ectothermic Vertebrates
by Jacques Robert and James K. Jancovich
Viruses 2016, 8(7), 187; https://doi.org/10.3390/v8070187 - 06 Jul 2016
Cited by 19 | Viewed by 7036
Abstract
Ranaviruses (Iridoviridae) are large DNA viruses that are causing emerging infectious diseases at an alarming rate in both wild and captive cold blood vertebrate species all over the world. Although the general biology of these viruses that presents some similarities with [...] Read more.
Ranaviruses (Iridoviridae) are large DNA viruses that are causing emerging infectious diseases at an alarming rate in both wild and captive cold blood vertebrate species all over the world. Although the general biology of these viruses that presents some similarities with poxvirus is characterized, many aspects of their replication cycles, host cell interactions and evolution still remain largely unclear, especially in vivo. Over several years, strategies to generate site-specific ranavirus recombinant, either expressing fluorescent reporter genes or deficient for particular viral genes, have been developed. We review here these strategies, the main ranavirus recombinants characterized and their usefulness for in vitro and in vivo studies. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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703 KiB  
Review
Newcastle Disease Virus as a Vaccine Vector for Development of Human and Veterinary Vaccines
by Shin-Hee Kim and Siba K. Samal
Viruses 2016, 8(7), 183; https://doi.org/10.3390/v8070183 - 04 Jul 2016
Cited by 86 | Viewed by 14091
Abstract
Viral vaccine vectors have shown to be effective in inducing a robust immune response against the vaccine antigen. Newcastle disease virus (NDV), an avian paramyxovirus, is a promising vaccine vector against human and veterinary pathogens. Avirulent NDV strains LaSota and B1 have long [...] Read more.
Viral vaccine vectors have shown to be effective in inducing a robust immune response against the vaccine antigen. Newcastle disease virus (NDV), an avian paramyxovirus, is a promising vaccine vector against human and veterinary pathogens. Avirulent NDV strains LaSota and B1 have long track records of safety and efficacy. Therefore, use of these strains as vaccine vectors is highly safe in avian and non-avian species. NDV replicates efficiently in the respiratory track of the host and induces strong local and systemic immune responses against the foreign antigen. As a vaccine vector, NDV can accommodate foreign sequences with a good degree of stability and as a RNA virus, there is limited possibility for recombination with host cell DNA. Using NDV as a vaccine vector in humans offers several advantages over other viral vaccine vectors. NDV is safe in humans due to host range restriction and there is no pre-existing antibody to NDV in the human population. NDV is antigenically distinct from common human pathogens. NDV replicates to high titer in a cell line acceptable for human vaccine development. Therefore, NDV is an attractive vaccine vector for human pathogens for which vaccines are currently not available. NDV is also an attractive vaccine vector for animal pathogens. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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1130 KiB  
Review
Replication-Competent Influenza A Viruses Expressing Reporter Genes
by Michael Breen, Aitor Nogales, Steven F. Baker and Luis Martínez-Sobrido
Viruses 2016, 8(7), 179; https://doi.org/10.3390/v8070179 - 23 Jun 2016
Cited by 48 | Viewed by 10293
Abstract
Influenza A viruses (IAV) cause annual seasonal human respiratory disease epidemics. In addition, IAV have been implicated in occasional pandemics with inordinate health and economic consequences. Studying IAV, in vitro or in vivo, requires the use of laborious secondary methodologies to identify virus-infected [...] Read more.
Influenza A viruses (IAV) cause annual seasonal human respiratory disease epidemics. In addition, IAV have been implicated in occasional pandemics with inordinate health and economic consequences. Studying IAV, in vitro or in vivo, requires the use of laborious secondary methodologies to identify virus-infected cells. To circumvent this requirement, replication-competent IAV expressing an easily traceable reporter protein can be used. Here we discuss the development and applications of recombinant replication-competent IAV harboring diverse fluorescent or bioluminescent reporter genes in different locations of the viral genome. These viruses have been employed for in vitro and in vivo studies, such as the screening of neutralizing antibodies or antiviral compounds, the identification of host factors involved in viral replication, cell tropism, the development of vaccines, or the assessment of viral infection dynamics. In summary, reporter-expressing, replicating-competent IAV represent a powerful tool for the study of IAV both in vitro and in vivo. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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1141 KiB  
Review
Marburg Virus Reverse Genetics Systems
by Kristina Maria Schmidt and Elke Mühlberger
Viruses 2016, 8(6), 178; https://doi.org/10.3390/v8060178 - 22 Jun 2016
Cited by 16 | Viewed by 10704
Abstract
The highly pathogenic Marburg virus (MARV) is a member of the Filoviridae family and belongs to the group of nonsegmented negative-strand RNA viruses. Reverse genetics systems established for MARV have been used to study various aspects of the viral replication cycle, analyze host [...] Read more.
The highly pathogenic Marburg virus (MARV) is a member of the Filoviridae family and belongs to the group of nonsegmented negative-strand RNA viruses. Reverse genetics systems established for MARV have been used to study various aspects of the viral replication cycle, analyze host responses, image viral infection, and screen for antivirals. This article provides an overview of the currently established MARV reverse genetic systems based on minigenomes, infectious virus-like particles and full-length clones, and the research that has been conducted using these systems. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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1730 KiB  
Review
Use of Reporter Genes in the Generation of Vaccinia Virus-Derived Vectors
by Sally Al Ali, Sara Baldanta, Mercedes Fernández-Escobar and Susana Guerra
Viruses 2016, 8(5), 134; https://doi.org/10.3390/v8050134 - 21 May 2016
Cited by 3 | Viewed by 12607
Abstract
Vaccinia virus (VACV) is one of the most extensively-studied viruses of the Poxviridae family. It is easy to genetically modify, so it has become a key tool for many applications. In this context, reporter genes facilitate the study of the role of foreign [...] Read more.
Vaccinia virus (VACV) is one of the most extensively-studied viruses of the Poxviridae family. It is easy to genetically modify, so it has become a key tool for many applications. In this context, reporter genes facilitate the study of the role of foreign genes introduced into the genome of VACV. In this review, we describe the type of reporter genes that have been used to generate reporter-expressing VACV and the applications of the recombinant viruses obtained. Reporter-expressing VACV are currently employed in basic and immunology research, in the development of vaccines and cancer treatment. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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704 KiB  
Review
Engineering Hepadnaviruses as Reporter-Expressing Vectors: Recent Progress and Future Perspectives
by Weiya Bai, Xiaoxian Cui, Youhua Xie and Jing Liu
Viruses 2016, 8(5), 125; https://doi.org/10.3390/v8050125 - 10 May 2016
Cited by 3 | Viewed by 4303
Abstract
The Hepadnaviridae family of small, enveloped DNA viruses are characterized by a strict host range and hepatocyte tropism. The prototype hepatitis B virus (HBV) is a major human pathogen and constitutes a public health problem, especially in high-incidence areas. Reporter-expressing recombinant viruses are [...] Read more.
The Hepadnaviridae family of small, enveloped DNA viruses are characterized by a strict host range and hepatocyte tropism. The prototype hepatitis B virus (HBV) is a major human pathogen and constitutes a public health problem, especially in high-incidence areas. Reporter-expressing recombinant viruses are powerful tools in both studies of basic virology and development of antiviral therapeutics. In addition, the highly restricted tropism of HBV for human hepatocytes makes it an ideal tool for hepatocyte-targeting in vivo applications such as liver-specific gene delivery. However, compact genome organization and complex replication mechanisms of hepadnaviruses have made it difficult to engineer replication-competent recombinant viruses that express biologically-relevant cargo genes. This review analyzes difficulties associated with recombinant hepadnavirus vector development, summarizes and compares the progress made in this field both historically and recently, and discusses future perspectives regarding both vector design and application. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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224 KiB  
Review
Applications of Replicating-Competent Reporter-Expressing Viruses in Diagnostic and Molecular Virology
by Yongfeng Li, Lian-Feng Li, Shaoxiong Yu, Xiao Wang, Lingkai Zhang, Jiahui Yu, Libao Xie, Weike Li, Razim Ali and Hua-Ji Qiu
Viruses 2016, 8(5), 127; https://doi.org/10.3390/v8050127 - 06 May 2016
Cited by 18 | Viewed by 5179
Abstract
Commonly used tests based on wild-type viruses, such as immunostaining, cannot meet the demands for rapid detection of viral replication, high-throughput screening for antivirals, as well as for tracking viral proteins or virus transport in real time. Notably, the development of replicating-competent reporter-expressing [...] Read more.
Commonly used tests based on wild-type viruses, such as immunostaining, cannot meet the demands for rapid detection of viral replication, high-throughput screening for antivirals, as well as for tracking viral proteins or virus transport in real time. Notably, the development of replicating-competent reporter-expressing viruses (RCREVs) has provided an excellent option to detect directly viral replication without the use of secondary labeling, which represents a significant advance in virology. This article reviews the applications of RCREVs in diagnostic and molecular virology, including rapid neutralization tests, high-throughput screening systems, identification of viral receptors and virus-host interactions, dynamics of viral infections in vitro and in vivo, vaccination approaches and others. However, there remain various challenges associated with RCREVs, including pathogenicity alterations due to the insertion of a reporter gene, instability or loss of the reporter gene expression, or attenuation of reporter signals in vivo. Despite all these limitations, RCREVs have become powerful tools for both basic and applied virology with the development of new technologies for generating RCREVs, the inventions of novel reporters and the better understanding of regulation of viral replication. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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Review
Dengue Virus Reporter Replicon is a Valuable Tool for Antiviral Drug Discovery and Analysis of Virus Replication Mechanisms
by Fumihiro Kato and Takayuki Hishiki
Viruses 2016, 8(5), 122; https://doi.org/10.3390/v8050122 - 05 May 2016
Cited by 32 | Viewed by 11952
Abstract
Dengue, the most prevalent arthropod-borne viral disease, is caused by the dengue virus (DENV), a member of the Flaviviridae family, and is a considerable public health threat in over 100 countries, with 2.5 billion people living in high-risk areas. However, no specific antiviral [...] Read more.
Dengue, the most prevalent arthropod-borne viral disease, is caused by the dengue virus (DENV), a member of the Flaviviridae family, and is a considerable public health threat in over 100 countries, with 2.5 billion people living in high-risk areas. However, no specific antiviral drug or licensed vaccine currently targets DENV infection. The replicon system has all the factors needed for viral replication in cells. Since the development of replicon systems, transient and stable reporter replicons, as well as reporter viruses, have been used in the study of various virological aspects of DENV and in the identification of DENV inhibitors. In this review, we summarize the DENV reporter replicon system and its applications in high-throughput screening (HTS) for identification of anti-DENV inhibitors. We also describe the use of this system in elucidation of the mechanisms of virus replication and viral dynamics in vivo and in vitro. Full article
(This article belongs to the Special Issue Replication-Competent Reporter-Expressing Viruses)
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