title: In vivo application of an RNAi strategy for the selective suppression of a mutant allele.
abstract: Gene therapy for dominantly inherited diseases with small interfering RNA (siRNA) requires mutant allele-specific suppression when genes in which mutation causes disease normally have an important role. We previously proposed a strategy for selective suppression of mutant alleles; both mutant and wild-type alleles are inhibited by most effective siRNA, and wild-type protein is restored using mRNA mutated to be resistant to the siRNA. Here, to prove the principle of this strategy in vivo, we applied it to our previously reported anti-copper/zinc superoxide dismutase (SOD1) short hairpin RNA (shRNA) transgenic (Tg) mice, in which the expression of the endogenous wild-type SOD1 gene was inhibited by more than 80%. These shRNA Tg mice showed hepatic lipid accumulation with mild liver dysfunction due to downregulation of endogenous wild-type SOD1. To rescue this side effect, we generated siRNA-resistant SOD1 Tg mice and crossed them with anti-SOD1 shRNA Tg mice, resulting in the disappearance of lipid accumulation in the liver. Furthermore, we also succeeded in mutant SOD1-specific gene suppression in the liver of SOD1(G93A) Tg mice, a model for amyotrophic lateral sclerosis, using intravenously administered viral vectors. Our method may prove useful for siRNA-based gene therapy for dominantly inherited diseases.
authors.affiliation: Department of Neurology and Neurological Science, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kubodera
authors.forename: Hiromi
authors.initials: H
authors.lastname: Yamada
authors.forename: Masayuki
authors.initials: M
authors.lastname: Anzai
authors.forename: Shinga
authors.initials: S
authors.lastname: Ohira
authors.forename: Shigefumi
authors.initials: S
authors.lastname: Yokota
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Hirai
authors.forename: Hideki
authors.initials: H
authors.lastname: Mochizuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.forename: Tasuku
authors.initials: T
authors.lastname: Mitani
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Mizusawa
authors.forename: Takanori
authors.initials: T
authors.lastname: Yokota
mesh_terms: D000483:Alleles; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism; D000818:Animals; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008099:Liver / Q000503:physiopathology*; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics*; D000072105:Superoxide Dismutase-1; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C000606290:SOD1 protein, human; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1