Infectious entry of reovirus cores into mammalian cells enhanced by transfection.
The reovirus virion is a moderately complex structure that contains eight structural proteins organized in multiple concentric capsid layers. Mammalian orthoreovirus virions undergo partial uncoating to produce infectious subvirion particles (ISVPs) and cores. Virions and ISVPs are infectious, whereas cores are transcriptionally active, but non-infectious, presumably because cores lack entry signals present in outer capsid proteins. We generated and purified reovirus cores and exposed them to cells with and without transfection reagents to directly test whether punfied cores can establish productive infections. Cores added directly to cells were essentially non-infectious. Specific infectivity of core/lipofectamine mixtures was increased more than 10,000-fold. Similar results were found with two reovirus serotypes (T1L and T3D) and in different cell types. These experiments indicated transfection reagents enhance core particle entry into cells and that cores contain all necessary components to replicate. This method may pave the way to improved genetic manipulations and to improved usage of reovirus as an anti-cancer agent.
Jiang
Jieyuan
J
Department of Medical Microbiology, University of Manitoba, 511 Basic Medical Sciences, 730 William Avenue, Winnipeg, Man., Canada R3N 0W3.
Coombs
Kevin M
KM
D000818:Animals; D002130:Calcium Phosphates; D002522:Chlorocebus aethiops; D007739:L Cells; D008055:Lipids; D012089:Mammalian orthoreovirus 3 / Q000235:genetics / Q000472:pathogenicity; D051379:Mice; D030102:Orthoreovirus, Mammalian / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D014162:Transfection; D014709:Vero Cells / Q000821:virology; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism*; D014773:Virology / Q000379:methods; D014779:Virus Replication
D002130:Calcium Phosphates; D008055:Lipids; C086724:Lipofectamine; D014758:Viral Core Proteins; C020243:calcium phosphate