Development of siRNA expression vector utilizing rock bream beta-actin promoter: a potential therapeutic tool against viral infection in fish.
In the present study, we have developed short interfering RNA (siRNA) expression vector utilizing rock bream beta-actin promoter and examined the possible use for the inhibition of highly pathogenic fish virus, rock bream iridovirus (RBIV), replication in vitro. Initially, in order to express siRNA effectively, we added several modifications to wild-type rock bream beta-actin promoter. Next, we succeeded in knocking down the expression of enhanced green fluorescent protein reporter gene expression in fish cells using newly developed vector more effectively than the fugu U6 promoter-driven vector we described previously. Finally, we could observe that cells transfected with modified rock bream beta-actin promoter-driven siRNA expression vector targeting major capsid protein (MCP) gene of RBIV exhibited more resistance to RBIV challenge than other control cells. Our results indicate that this novel siRNA expression vector can be used as a new tool for therapeutics in virus infection in fish species.
Zenke
Kosuke
K
Department of Aquatic Life Medicine, Pukyong National University, Nam-gu 599-1, Busan, South Korea.
Nam
Yoon Kwon
YK
Kim
Ki Hong
KH
D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D005393:Fish Diseases / Q000628:therapy*; D005399:Fishes; D055785:Gene Knockdown Techniques; D015316:Genetic Therapy / Q000379:methods*; D017994:Iridovirus / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014777:Virus Diseases / Q000628:therapy*
D000199:Actins; D034741:RNA, Small Interfering