title: Allele-specific silencing of dominant disease genes.
abstract: Small interfering RNA (siRNA) holds therapeutic promise for silencing dominantly acting disease genes, particularly if mutant alleles can be targeted selectively. In mammalian cell models we demonstrate that allele-specific silencing of disease genes with siRNA can be achieved by targeting either a linked single-nucleotide polymorphism (SNP) or the disease mutation directly. For a polyglutamine neurodegenerative disorder in which we first determined that selective targeting of the disease-causing CAG repeat is not possible, we took advantage of an associated SNP to generate siRNA that exclusively silenced the mutant Machado-Joseph disease/spinocerebellar ataxia type 3 allele while sparing expression of the WT allele. Allele-specific suppression was accomplished with all three approaches currently used to deliver siRNA: in vitro-synthesized duplexes as well as plasmid and viral expression of short hairpin RNA. We further optimized siRNA to specifically target a missense Tau mutation, V337M, that causes frontotemporal dementia. These studies establish that siRNA can be engineered to silence disease genes differing by a single nucleotide and highlight a key role for SNPs in extending the utility of siRNA in dominantly inherited disorders.
authors.affiliation: Department of Neurology, Graduate Program in Genetics, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA 52242, USA.
authors.forename: Victor M
authors.initials: VM
authors.lastname: Miller
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
authors.forename: Ginger L
authors.initials: GL
authors.lastname: Marrs
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Gouvion
authors.forename: Gloria
authors.initials: G
authors.lastname: Lee
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
authors.forename: Henry L
authors.initials: HL
authors.lastname: Paulson
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003704:Dementia / Q000235:genetics / Q000628:therapy; D020868:Gene Silencing; D005799:Genes, Dominant; D030342:Genetic Diseases, Inborn / Q000235:genetics* / Q000628:therapy*; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D017827:Machado-Joseph Disease / Q000235:genetics / Q000628:therapy; D010455:Peptides / Q000235:genetics; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D034741:RNA, Small Interfering / Q000235:genetics* / Q000494:pharmacology*; D016875:tau Proteins / Q000235:genetics
chemical_list: D010455:Peptides; D034741:RNA, Small Interfering; D016875:tau Proteins; C097188:polyglutamine