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

Piyasena, Thisun B. H. , Setoh, Yin X., Hobson-Peters, Jody, Newton, Natalee D. , Bielefeldt-Ohmann, Helle, McLean, Breeanna J. , Vet, Laura J. , Khromykh, Alexander A. and Hall, Roy A. (2017) Infectious DNAs derived from insect-specific flavivirus genomes enable identification of pre- and post-entry host restrictions in vertebrate cells. Scientific Reports, 7 1: 2940.1-2940.11. doi:10.1038/s41598-017-03120-1


Author Piyasena, Thisun B. H.
Setoh, Yin X.
Hobson-Peters, Jody
Newton, Natalee D.
Bielefeldt-Ohmann, Helle
McLean, Breeanna J.
Vet, Laura J.
Khromykh, Alexander A.
Hall, Roy A.
Title Infectious DNAs derived from insect-specific flavivirus genomes enable identification of pre- and post-entry host restrictions in vertebrate cells
Journal name Scientific Reports   Check publisher's open access policy
ISSN 2045-2322
Publication date 2017-06-07
Year available 2017
Sub-type Article (original research)
DOI 10.1038/s41598-017-03120-1
Open Access Status DOI
Volume 7
Issue 1
Start page 2940.1
End page 2940.11
Total pages 11
Place of publication London, United Kingdom
Publisher Nature Publishing Group
Language eng
Abstract Flaviviruses such as West Nile virus (WNV), dengue virus and Zika virus are mosquito-borne pathogens that cause significant human diseases. A novel group of insect-specific flaviviruses (ISFs), which only replicate in mosquitoes, have also been identified. However, little is known about the mechanisms of ISF host restriction. We report the generation of infectious cDNA from two Australian ISFs, Parramatta River virus (PaRV) and Palm Creek virus (PCV). Using circular polymerase extension cloning (CPEC) with a modified OpIE2 insect promoter, infectious cDNA was generated and transfected directly into mosquito cells to produce infectious virus indistinguishable from wild-type virus. When infectious PaRV cDNA under transcriptional control of a mammalian promoter was used to transfect mouse embryo fibroblasts, the virus failed to initiate replication even when cell entry steps were by-passed and the type I interferon response was lacking. We also used CPEC to generate viable chimeric viruses between PCV and WNV. Analysis of these hybrid viruses revealed that ISFs are also restricted from replication in vertebrate cells at the point of entry. The approaches described here to generate infectious ISF DNAs and chimeric viruses provide unique tools to further dissect the mechanisms of their host restriction.
Keyword Flaviviruses
West Nile virus (WNV)
Dengue virus
Zika virus
Q-Index Code C1
Grant ID ARC DP120103994
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
School of Chemistry and Molecular Biosciences
 
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Created: Fri, 16 Jun 2017, 12:25:15 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences