Packaging the replicon RNA of the Far-Eastern subtype of tick-borne encephalitis virus into single-round infectious particles: development of a heterologous gene delivery system.
The sub-genomic replicon of tick-borne encephalitis (TBE) virus (Far-Eastern subtype) was packaged into infectious particles by providing the viral structural proteins in trans. Sequential transfection of TBE replicon RNA and a plasmid that expressed the structural proteins led to the secretion of infectious particles that contained TBE replicon RNA. The secreted particles had single-round infectivity, which was inhibited by TBE virus-neutralizing antibody. The physical structure of the particles was almost identical to that of infectious virions, and the packaged replicon RNA showed no recombination with the mRNAs of the viral structural proteins. Furthermore, heterologous genes were successfully delivered and expressed by packaging TBE replicon RNA with inserted GFP and Neo genes. This replicon packaging system may be a useful tool for the molecular study of the TBE virus genome packaging mechanism, and for the development of vaccine delivery systems.
Yoshii
Kentarou
K
Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Kita-18 Nishi-9, Kita-ku, Sapporo, Hokkaido 060-0818, Japan.
Hayasaka
Daisuke
D
Goto
Akiko
A
Kawakami
Kazue
K
Kariwa
Hiroaki
H
Takashima
Ikuo
I
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics* / Q000472:pathogenicity; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D010957:Plasmids / Q000276:immunology; D012367:RNA, Viral / Q000235:genetics* / Q000276:immunology; D012093:Replicon / Q000235:genetics* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D000914:Antibodies, Viral; D012367:RNA, Viral; D014764:Viral Proteins