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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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.
Commonly used reporter...

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Methodology for the Efficient Generation of Fluorescently Tagged Vaccinia Virus Proteins
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Fluorescent protein-expressing Modified Vaccinia Ankara encoding T7 RNA polymerase.

Nathaniel Jackson1, Mahmoud Bayoumi1,2, Himadri Nath1

  • 1Host-pathogen interactions and Disease Intervention and Prevention programs, Texas Biomedical Research Institute, San Antonio, Texas, USA.

Journal of Virology
|June 12, 2026
PubMed
Summary

We developed fluorescent Modified Vaccinia Ankara-T7 (MVA-T7) viruses for improved recovery of negative-stranded RNA viruses. These fluorescent MVA-T7 viruses enable direct visualization of infection and simplify virus stock management, enhancing reverse genetics efficiency.

Keywords:
MVARABVT7 RNA polymeraseVSVreporter virusreverse genetics

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Area of Science:

  • Virology
  • Molecular Biology
  • Gene Expression Systems

Background:

  • Efficient recovery of negative-stranded RNA viruses relies on bacteriophage T7 RNA polymerase, often supplied by Modified Vaccinia Ankara expressing T7 polymerase (MVA T7).
  • Challenges exist in generating virus stocks, monitoring MVA T7 infection, and detecting residual helper virus during recombinant virus rescue.

Purpose of the Study:

  • To generate and characterize fluorescent protein-expressing MVA T7 viruses (EGFP and mRFP1) for improved visualization and management in reverse genetics.
  • To assess if fluorescent MVA T7 viruses maintain growth kinetics and transgene expression comparable to parental MVA T7.
  • To evaluate the utility of fluorescent MVA T7 viruses in facilitating recombinant virus rescue and detecting helper virus carryover.

Main Methods:

  • Generation of MVA T7 viruses expressing enhanced green fluorescent protein (EGFP) or monomeric red fluorescent protein 1 (mRFP1).
  • Characterization of fluorescent MVA T7 viruses for plaque morphology, growth kinetics, and transgene expression.
  • Assessment of transient protein expression and rescue of recombinant vesicular stomatitis virus (rVSV) using fluorescent MVA T7 viruses.
  • Evaluation of helper virus detection during rVSV recovery and amplification.

Main Results:

  • Fluorescent MVA T7 viruses (EGFP and mRFP1) exhibited comparable plaque morphology, growth kinetics, and transgene expression to parental MVA T7.
  • Fluorescent MVA T7 viruses efficiently supported foreign gene expression and rVSV rescue.
  • Direct visualization of MVA T7 infection was enabled by fluorescent proteins.
  • Fluorescent MVA T7 viruses facilitated stock generation, titration, and detection of helper virus carryover.

Conclusions:

  • EGFP- and mRFP1-expressing MVA T7 viruses are practical tools for monitoring T7-driven rescue of recombinant RNA viruses.
  • These fluorescent viruses improve the efficiency, reproducibility, and quality control of reverse genetics systems.
  • They simplify stock management and enable rapid identification of helper virus contamination, enhancing viral rescue and amplification processes.