Multi-color fluorescent reporter dengue viruses with improved stability for analysis of a multi-virus infection

PLoS One. 2018 Mar 16;13(3):e0194399. doi: 10.1371/journal.pone.0194399. eCollection 2018.

Abstract

Reporter virus is a versatile tool to visualize and to analyze virus infections. However, for flaviviruses, it is difficult to maintain the inserted reporter genes on the viral genome, limiting its use in several studies that require homogeneous virus particles and several rounds of virus replication. Here, we showed that flanking inserted GFP genes on both sides with ribosome-skipping 2A sequences improved the stability and the consistency of their fluorescent signals for dengue-virus-serotype 2 (DENV2) reporter viruses. The reporter viruses can infect known susceptible mammalian cell lines and primary CD14+ human monocytes. This design can accommodate several fluorescent protein genes, enabling the generation of multi-color DENV2-16681 reporter viruses with comparable replication capabilities, as demonstrated by their abilities to maintain their fluorescent intensities during co-infections and to exclude superinfections regardless of the fluorescent tags. The reported design of multi-color DENV2 should be useful for high-throughput analyses, single-cell analysis, and characterizations of interference and superinfection in animal models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cells, Cultured
  • Chlorocebus aethiops
  • Dengue Virus / genetics*
  • Dengue Virus / metabolism
  • Genes, Reporter / genetics
  • Genome, Viral / genetics*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • K562 Cells
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence
  • Vero Cells
  • Virus Replication / genetics*

Substances

  • Luminescent Proteins
  • Green Fluorescent Proteins

Grants and funding

This work is funded by a grant from Grand Challenges Canada, Stars in Global Health Program Round 5 to BS.