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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Production of OSU G5P[7] Porcine Rotavirus Expressing a Fluorescent Reporter via Reverse Genetics.

Anthony J Snyder1, Chantal A Agbemabiese1,2, John T Patton1

  • 1Department of Biology, Indiana University, 212 S. Hawthorne Drive, Simon Hall 011, Bloomington, IN 47405, USA.

Viruses
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Researchers developed a new reverse genetics system for porcine rotavirus, creating a modified virus that can express foreign proteins. This innovation could lead to improved vaccines against rotavirus gastroenteritis in piglets.

Keywords:
expression vectorporcine rotavirusreverse geneticsrotavirus

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Area of Science:

  • Veterinary Virology
  • Molecular Biology
  • Vaccine Development

Background:

  • Porcine rotavirus causes severe gastroenteritis in piglets, necessitating improved vaccines.
  • Current porcine rotavirus vaccines exhibit suboptimal efficacy.
  • The G5P[7] genotype of Rotavirus A is prevalent in piglet disease.

Purpose of the Study:

  • To develop a robust reverse genetics system for the OSU rotavirus strain (G5P[7]).
  • To engineer a recombinant porcine rotavirus capable of expressing foreign proteins.
  • To explore the potential for novel vaccine strategies against porcine rotavirus.

Main Methods:

  • Complete genome sequencing of the OSU rotavirus strain using Nanopore technology.
  • Establishment of a reverse genetics system for generating recombinant (r)OSU rotavirus.
  • Engineering of the rOSU genome to express the fluorescent UnaG protein via a fused NSP3-2A-UnaG open reading frame.

Main Results:

  • A functional reverse genetics system was successfully established for the OSU rotavirus.
  • Recombinant OSU rotavirus (rOSU) was produced with high titers (~10^7 PFU/mL), albeit with slightly reduced growth kinetics.
  • Genetically stable rOSU expressing functional, separate UnaG protein was generated, demonstrating its utility as an expression vector.

Conclusions:

  • The developed reverse genetics system enables the creation of genetically modified porcine rotaviruses.
  • Recombinant OSU rotavirus can serve as a platform for expressing foreign antigens.
  • This technology holds promise for developing next-generation porcine rotavirus vaccines, including combination vaccines.