Allele-Specific Silencing of Dominant Disease Genes

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...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2003-06, Vol.100 (12), p.7195-7200
Main Authors: Miller, Victor M., Xia, Haibin, Marrs, Ginger L., Gouvion, Cynthia M., Lee, Gloria, Davidson, Beverly L., Paulson, Henry L.
Format: Article
Language:English
Subjects:
Alleles
Animals
Base Sequence
Biological Sciences
Cell lines
COS Cells
Dementia - genetics
Dementia - therapy
Disease
Fluorescence
Gene expression
Gene Silencing
Genes, Dominant
Genetic diseases
Genetic Diseases, Inborn - genetics
Genetic Diseases, Inborn - therapy
Genetic Therapy
Genetics
HeLa Cells
Humans
Machado-Joseph Disease - genetics
Machado-Joseph Disease - therapy
Nervous system diseases
Neurodegenerative diseases
Peptides - genetics
Plasmids
Point Mutation
Polymorphism, Single Nucleotide
Ribonucleic acid
RNA
RNA, Small Interfering - genetics
RNA, Small Interfering - pharmacology
Small interfering RNA
tau Proteins - genetics
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1231012100