Selective silencing by RNAi of a dominant allele that causes amyotrophic lateral sclerosis
Schwarz, Dianne S.
Keene, Alex Carl
Affar, El Bachir
Zamore, Phillip D.
Student AuthorsAlex Keene
UMass Chan AffiliationsDepartment of Biochemistry and Molecular Pharmacology
Document TypeJournal Article
KeywordsAlleles; 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
Medicine and Health Sciences
Neuroscience and Neurobiology
MetadataShow full item record
AbstractRNA 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.
Aging Cell. 2003 Aug;2(4):209-17.
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/33646