Functional modification of Sendai virus by siRNA.
Sendai virus (hemagglutinating virus of Japan; HVJ) is a negative-strand RNA virus with robust fusion activity, and has been utilized for gene transfer and drug delivery. Hemagglutinin-neuraminidase (HN) protein on the viral membrane is important for cell fusion, but causes agglutination of red blood cells. HN-depleted HVJ has been desired for in vivo transfection in order to improve safety. Here, we succeeded in producing HN-depleted HVJ using HN-specific short interfering RNA (siRNA). Viral production was not affected by the siRNA. HN protein was markedly decreased in the new HVJ, while other viral proteins were retained. Consequently, the hemagglutinating activity was substantially reduced and infection activity was suppressed. When the HN-depleted HVJ was mixed with cultured cells and the mixture was centrifuged for 10min at 2000xg, the modified HVJ recovered its infectivity to approximately 80% of wild HVJ. However, infectivity was abolished in the presence of anti-F antibody. Moreover, transfection of FITC-labeled oligodeoxynucleotides using the modified HVJ was also recovered by centrifugation. Thus, the HN-depleted HVJ produced using siRNA technology will be applicable to a delivery vector.
Saga
Kotaro
K
Division of Gene Therapy Science, Graduate School of Medicine, Osaka University, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Tamai
Katsuto
K
Kawachi
Masako
M
Shimbo
Takashi
T
Fujita
Hiroshi
H
Yamazaki
Takehiko
T
Kaneda
Yasufumi
Y
D000818:Animals; D002460:Cell Line; D002645:Chickens; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D015707:HN Protein / Q000378:metabolism; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D010253:Respirovirus Infections; D029082:Sendai virus / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection; D053586:Virus Internalization
D015707:HN Protein; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016650:Fluorescein-5-isothiocyanate