title: Development of an improved polykaryon-based influenza virus rescue system.
abstract: BACKGROUND
  Virus rescue from transfected cells is an extremely useful technique that allows defined viral clones to be engineered for the purpose of rational vaccine design or fundamental reverse genetics studies. However, it is often hindered by low primary rescue success rates or yields, especially with field-derived viral strains.
  
  
  METHODS
  We investigated the possibility of enhancing influenza virus rescue by eliciting cell fusion to increase the chances of having all necessary plasmids expressed within the same polykaryon. To this end we used the Maedi-Visna Virus envelope protein which has potent fusion activity in cells from a wide range of different species.
  
  
  RESULTS
  Co-transfecting cells with the eight plasmids necessary to rescue influenza virus plus a plasmid expressing the Maedi-Visna Virus envelope protein resulted in increased rescue efficiency. In addition, partial complements of the 8-plasmid rescue system could be transfected into two separate populations of cells, which upon fusion led to live virus rescue.
  
  
  CONCLUSIONS
  The simple modification described here has the potential to improve the efficiency of the virus rescue process and expand the potential applications for reverse genetic studies.
authors.affiliation: Cambridge Infectious Disease Consortium, Department of Veterinary Medicine, University of Cambridge, Cambridge, UK. v.bourret@envt.fr
authors.forename: Vincent
authors.initials: V
authors.lastname: Bourret
authors.forename: Jon
authors.initials: J
authors.lastname: Lyall
authors.forename: Mariette F
authors.initials: MF
authors.lastname: Ducatez
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Guérin
authors.forename: Laurence
authors.initials: L
authors.lastname: Tiley
mesh_terms: D003001:Cloning, Molecular; D057809:HEK293 Cells; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification; D009975:Orthomyxoviridae / Q000235:genetics / Q000302:isolation & purification*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014790:Visna-maedi virus / Q000378:metabolism
chemical_list: D012367:RNA, Viral; D014759:Viral Envelope Proteins