Selective silencing by RNAi of a dominant allele that causes amyotrophic lateral sclerosis
Department of Biochemistry and Molecular Pharmacology
Medical Subject Headings
Alleles; Amyotrophic Lateral Sclerosis; Animals; Drosophila melanogaster; Gene Expression Regulation, Enzymologic; Gene Silencing; Gene Therapy; Genes, Dominant; Genetic Vectors; Green Fluorescent Proteins; Humans; Luminescent Proteins; Mice; Point Mutation; RNA Interference; RNA Polymerase III; RNA, Small Interfering; Superoxide Dismutase; Transfection
Life Sciences | Medicine and Health Sciences | Neuroscience and Neurobiology
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.
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Citation: Aging Cell. 2003 Aug;2(4):209-17.