Recombinant Rotavirus
Recombinant Rotavirus - Related Videos
03:43
Plasmid-Mediated Generation of Recombinant Rotavirus in Cultured Cells
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2026
Source: Philip, A. A., et al. Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins. J. Vis. Exp. (2020)This video demonstrates the recovery of recombinant rotavirus by transfecting plasmids encoding viral genome segments into hamster kidney cells, followed by amplification in monkey epithelial cells, enabling studies of rotavirus biology, replication, and vaccine development.
04:12
Visualizing Trypsin-Activated Rotavirus Infection Using a Plaque Assay
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2026
Source: Philip, A. A., et. al., Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins. J. Vis. Exp. (2020)This video demonstrates a plaque assay technique to monitor rotavirus infection in epithelial cells. Trypsin activation enhances viral entry, and staining highlights viable cells, enabling visualization of infection-induced plaques.
03:44
Isolation and Visualization of Rotavirus Genome via Gel Electrophoresis
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2026
Source: Philip, A. A., et al., Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins. J. Vis. Exp. (2020)This video demonstrates the isolation of rotavirus double-stranded RNA from clarified cell lysates. It further illustrates the separation and visualization of genome segments through polyacrylamide gel electrophoresis.
03:30
Assessing Rotavirus Growth Curve by Plaque Assay
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2026
Source: Kadoya, S., Sano, D. Assays for the Specific Growth Rate and Cell-binding Ability of Rotavirus. J. Vis. Exp. (2019).This video demonstrates the use of a plaque assay to assess the rotavirus growth curve. The method involves infecting host cells with diluted virus, overlaying with agarose to restrict spread, and staining to visualize plaques formed by viral replication. By counting plaque-forming units at defined time points, the resulting growth curve reveals the lag, exponential, and...
11:40
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Cited by 24 •
2020
Generation of recombinant rotaviruses from plasmid DNA provides an essential tool for the study of rotavirus replication and pathogenesis, and the development of rotavirus expression vectors and vaccines. Herein, we describe a simplified reverse genetics approach for generating recombinant rotaviruses, including strains expressing fluorescent reporter proteins.
FAQs
Its segmented double-stranded RNA genome provides separate genetic units that can be targeted individually or in defined combinations. Reverse-genetics systems use this organization to introduce specified sequence changes and then recover virus in cultured cells. This controlled design lets investigators connect a particular viral protein or mutation with effects on entry, replication, disease-related processes, or host immune interactions.
Defined mutations allow researchers to examine the consequences of changing a specific viral feature rather than relying on uncontrolled genetic differences. Comparing engineered viruses with different alterations can clarify the contribution of particular proteins or genome segments to viral behavior. This approach helps connect molecular changes with processes such as cell entry, replication, disease development, and immune interaction.
Researchers can plan experiments around individual genome segments rather than treating viral genetic information as one undifferentiated unit. That arrangement supports targeted modification of selected segments and comparison of viruses carrying different engineered combinations. Such comparisons help attribute observed changes to particular genetic components, strengthening analysis of protein function, replication behavior, or interactions with host immunity.
A typical workflow begins by selecting one or more viral genome segments for modification and defining the intended genetic change. Researchers then use a reverse-genetics system to introduce those changes and attempt virus recovery in cultured cells. The resulting virus can be examined in controlled experiments to assess its biological properties or its suitability for vaccine and delivery applications.
Cultured cells provide the setting in which researchers can recover engineered virus after applying a reverse-genetics system. They also create a controlled experimental context for examining how a selected mutation or viral protein affects rotavirus behavior. This recovery step makes it possible to evaluate engineered candidates systematically before considering their use in studies of immunity, vaccination, antiviral strategies, or delivery technologies.
These engineered viruses support evaluation of attenuated vaccine candidates and investigation of antiviral strategies. Because researchers can introduce defined genetic changes, they can examine how those changes relate to viral biology and host immune responses in a controlled setting. The resulting observations help identify candidate designs or viral features that warrant further study in vaccine development and antiviral research.
Engineered rotavirus platforms can be investigated for presenting or delivering selected antigens. In this context, genetic modification is used not only to study the virus itself but also to explore how it might carry biologically relevant antigenic material. These applications extend recombinant rotavirus research beyond basic biology toward vaccine technologies and experimental delivery systems.