title: Selective silencing by RNAi of a dominant allele that causes amyotrophic lateral sclerosis.
abstract: RNA interference (RNAi) can achieve sequence-selective inactivation of gene expression in a wide variety of eukaryotes by introducing double-stranded RNA corresponding to the target gene. Here we explore the potential of RNAi as a therapy for amyotrophic lateral sclerosis (ALS) caused by mutations in the Cu, Zn superoxide dismutase (SOD1) gene. Although the mutant SOD1 is toxic, the wild-type SOD1 performs important functions. Therefore, the ideal therapeutic strategy should be to selectively inhibit the mutant, but not the wild-type SOD1 expression. Because most SOD1 mutations are single nucleotide changes, to selectively silence the mutant requires single-nucleotide specificity. By coupling rational design of small interfering RNAs (siRNAs) with their validation in RNAi reactions in vitro and in vivo, we have identified siRNA sequences with this specificity. A similarly designed sequence, when expressed as small hairpin RNA (shRNA) under the control of an RNA polymerase III (pol III) promoter, retains the single-nucleotide specificity. Thus, RNAi is a promising therapy for ALS and other disorders caused by dominant, gain-of-function gene mutations.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605-2324, USA.
authors.forename: Hongliu
authors.initials: H
authors.lastname: Ding
authors.forename: Dianne S
authors.initials: DS
authors.lastname: Schwarz
authors.forename: Alex
authors.initials: A
authors.lastname: Keene
authors.forename: El Bachir
authors.initials: el B
authors.lastname: Affar
authors.forename: Laura
authors.initials: L
authors.lastname: Fenton
authors.forename: Xugang
authors.initials: X
authors.lastname: Xia
authors.forename: Yang
authors.initials: Y
authors.lastname: Shi
authors.forename: Phillip D
authors.initials: PD
authors.lastname: Zamore
authors.forename: Zuoshang
authors.initials: Z
authors.lastname: Xu
mesh_terms: D000483:Alleles; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000628:therapy*; D000818:Animals; D004331:Drosophila melanogaster; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D020868:Gene Silencing / Q000502:physiology*; D005799:Genes, Dominant / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D051379:Mice; D017354:Point Mutation / Q000235:genetics; D034622:RNA Interference / Q000502:physiology*; D012320:RNA Polymerase III / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000172:deficiency / Q000235:genetics*; D000072105:Superoxide Dismutase-1; D014162:Transfection
chemical_list: D008164:Luminescent Proteins; D034741:RNA, Small Interfering; C000606290:SOD1 protein, human; D049452:Green Fluorescent Proteins; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D012320:RNA Polymerase III