title: Multiple shRNAs driven by U6 and CMV promoter enhances efficiency of antiviral effects against foot-and-mouth disease virus.
abstract: Foot-and-mouth disease (FMD) is an economically significant animal disease because of the speed of its transmission. The current vaccine for FMD provides no protection until 7 days post-vaccination, thus reducing its effectiveness in the case of an outbreak. Small interfering RNA (siRNA) is a promising antiviral approach because it can induce a protective response much more rapidly. This study is the first report to apply multiple short hairpin RNA (shRNA) expression systems to inhibit foot-and-mouth disease virus (FMDV) replication. Three different shRNAs, one targeting 2B region and two targeting 3C region, were driven by three RNA Polymerase III (Pol III) promoters, U6 or a combination of two U6 promoters and one RNA Polymerase II (Pol II) promoter, CMV. The adenoviruses simultaneously expressing three different shRNAs in a single construct had significantly enhanced antiviral effects compared with those expressing only a single shRNA, those expressing double shRNAs or a mixture of adenoviruses expressing a single shRNA and the adenovirus expressing double shRNAs, both in vitro and in vivo. The adenoviruses had broad antiviral effects against seven serotypes of FMDV, including O, A, Asia1, C, SAT1, SAT2, and SAT3 in vitro, but differed in their efficacy. The adenovirus expressing multiple shRNAs driven by three U6 promoters had strong antiviral effects in suckling mice challenged with O, A, and Asia1 serotype of FMDV.
authors.affiliation: Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, Ministry for Food, Agriculture, Forestry and Fisheries, Anyang 430-824, Gyeonggi-do, Republic of Korea.
authors.forename: Su-Mi
authors.initials: SM
authors.lastname: Kim
authors.forename: Kwang-Nyeong
authors.initials: KN
authors.lastname: Lee
authors.forename: Su-Jung
authors.initials: SJ
authors.lastname: Lee
authors.forename: Young-Joon
authors.initials: YJ
authors.lastname: Ko
authors.forename: Hyang-Sim
authors.initials: HS
authors.lastname: Lee
authors.forename: Chang-Hee
authors.initials: CH
authors.lastname: Kweon
authors.forename: Hyun-Soo
authors.initials: HS
authors.lastname: Kim
authors.forename: Jong-Hyeon
authors.initials: JH
authors.lastname: Park
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D000998:Antiviral Agents / Q000494:pharmacology*; D001688:Biological Products / Q000096:biosynthesis / Q000494:pharmacology*; D003587:Cytomegalovirus / Q000235:genetics; D004195:Disease Models, Animal; D005536:Foot-and-Mouth Disease / Q000628:therapy; D029544:Foot-and-Mouth Disease Virus / Q000187:drug effects* / Q000472:pathogenicity*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D051379:Mice; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000096:biosynthesis / Q000494:pharmacology*; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D001688:Biological Products; D034741:RNA, Small Interfering