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Infectious DNAs derived from insect-specific flavivirus genomes enable identification of pre- and post-entry host

Thisun B H Piyasena1, Yin X Setoh1, Jody Hobson-Peters1

  • 1Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, 4072, Queensland, Australia.

Scientific Reports
|June 9, 2017
PubMed

Insights

Insect-specific flaviviruses (ISFs) like Parramatta River virus (PaRV) and Palm Creek virus (PCV) cannot replicate in vertebrate cells. Research shows ISFs are blocked at the entry stage, offering new tools to study host restriction mechanisms.

Area of Science:

  • Virology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Flaviviruses, including West Nile virus (WNV), dengue virus, and Zika virus, are significant mosquito-borne human pathogens.
  • Insect-specific flaviviruses (ISFs) are a distinct group that replicate only in mosquitoes, but their host restriction mechanisms remain poorly understood.

Purpose of the Study:

  • To generate infectious complementary DNA (cDNA) from Australian insect-specific flaviviruses (ISFs) Parramatta River virus (PaRV) and Palm Creek virus (PCV).
  • To investigate the mechanisms of host restriction for ISFs in vertebrate cells.
  • To develop novel tools for studying ISF-mammalian interactions.

Main Methods:

  • Infectious cDNA of PaRV and PCV was generated using circular polymerase extension cloning (CPEC) with a modified OpIE2 insect promoter.
  • Infectious virus was produced by transfecting mosquito cells with the generated cDNA.
  • Chimeric viruses were created between PCV and WNV using CPEC.
  • Replication competence of PaRV in mouse embryo fibroblasts was assessed using mammalian promoters, bypassing cellular entry and interferon responses.

Main Results:

  • Infectious PaRV and PCV were successfully generated from cDNA and produced infectious virus in mosquito cells.
  • PaRV failed to replicate in mouse embryo fibroblasts even when cellular entry and type I interferon responses were circumvented.
  • Analysis of chimeric viruses indicated that ISFs are restricted from replicating in vertebrate cells at the point of entry.

Conclusions:

  • Insect-specific flaviviruses (ISFs) are blocked from replicating in vertebrate cells, specifically at the entry stage.
  • The generation of infectious ISF cDNAs and chimeric viruses provides valuable tools for further research into ISF host restriction mechanisms.
  • Understanding ISF host restriction can inform strategies against related flaviviruses impacting human health.

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