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Functional modification of Sendai virus by siRNA

Author:
Saga, Kotaro, Tamai, Katsuto, Kawachi, Masako, Shimbo, Takashi, Fujita, Hiroshi, Yamazaki, Takehiko, Kaneda, Yasufumi
Source:
Journal of biotechnology 2008 v.133 no.3 pp. 386-394
ISSN:
0168-1656
Subject:
Sendai virus, antibodies, biotechnology, cell fusion, centrifugation, cultured cells, drugs, erythrocytes, hemagglutination, oligodeoxyribonucleotides, pathogenicity, small interfering RNA, transfection, viral proteins, viruses, Japan
Abstract:
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 2000x g, 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.
Agid:
274188