siRNA-based inhibition specific for mutant SOD1 with single nucleotide alternation in familial ALS, compared with ribozyme and DNA enzyme.
In many of autosomal dominant diseases such as familial amyotrophic lateral sclerosis (ALS) with SOD1 mutation, a missense point mutation may induce the disease by its gain of adverse property. Reduction of such a mutant protein expression is expected to improve the disease phenotype. Duplex of 21-nt RNA, known as siRNA, has recently emerged as a powerful tool to silence gene, but the sequence specificity and efficacies have not been fully studied in comparison with ribozyme and DNA enzyme. We could make the siRNA which recognized even a single nucleotide alternation and selectively suppress G93A SOD1 expression leaving wild-type SOD1 intact. In mammalian cells, the siRNA much more efficiently suppressed the expression of mutant SOD1 than ribozyme or DNA enzyme. Furthermore, these siRNAs could suppress cell death of Neuro2a induced by over-expression of mutant SOD1s with stress of proteasome inhibition. Our results support the feasibility of utilizing siRNA-based gene therapy of familial ALS with mutant SOD1.
Yokota
Takanori
T
Department of Neurology, Tokyo Medical and Dental University, Tokyo, Japan. tak-yokota.nuro@tmd.ac.jp
Miyagishi
Makoto
M
Hino
Taro
T
Matsumura
Ryusuke
R
Tasinato
Andrea
A
Urushitani
Makoto
M
Rao
Rammohan V
RV
Takahashi
Ryosuke
R
Bredesen
Dale E
DE
Taira
Kazunari
K
Mizusawa
Hidehiro
H
Andrea
Tasinato
T
D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D000818:Animals; D002460:Cell Line; D021881:DNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D005240:Feasibility Studies; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000378:metabolism*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009447:Neuroblastoma / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism*; D000072105:Superoxide Dismutase-1
D021881:DNA, Catalytic; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; C000606290:SOD1 protein, human; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1