Preferential silencing of a common dominant rhodopsin mutation does not inhibit retinal degeneration in a transgenic model

被引:33
作者
Tessitore, Alessandra
Parisi, Fabiana
Denti, Michela Alessandra
Allocca, Mariacarmela
Di Vicino, Umberto
Domenici, Luciano
Bozzoni, Irene
Auricchio, Alberto
机构
[1] Telethon Inst Genet & Med, I-80131 Naples, Italy
[2] Univ Roma La Sapienza, Dept Genet & Mol Biol, Inst Pasteur Cenci Bolognetti, Rome, Italy
[3] CNR, IBPM, Rome, Italy
[4] European Sch Mol Med, SEMM, Naples Site, Italy
[5] CNR, Inst Neurosci, I-56100 Pisa, Italy
[6] Univ Aquila, Dept Biomed Sci & Technol, I-67010 Laquila, Italy
[7] Univ Naples Federico II, Dept Pediat, Naples, Italy
关键词
retinitis pigmentosa; rhodopsin; P23H; photoreceptor cells; RNA interference; AAV;
D O I
10.1016/j.ymthe.2006.07.008
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Autosomal dominant retinitis pigmentosa caused by the frequent rhodopsin P23H mutation is characterized by progressive photoreceptor cell death eventually leading to blindness and for which no therapies are available. Considering the gain-of-function effect exerted by the P23H mutation, strategies aimed at silencing the expression of the mutated allele, like RNA interference, are desirable. We have designed small interfering RNAs (siRNA) to silence specifically the P23H rhodopsin allele expressed by a transgenic rat model of the disease. We have selected in vitro one siRNA and generated an adeno-associated viral (AAV) vector expressing the short hairpin RNA (shRNA) based on the selected siRNA. In vitro the shRNA significantly inhibits the expression of the P23H but not the wild-type rhodopsin allele. Subretinal administration of the AAV2/5 vector encoding the shRNA in P23H transgenic rats results in inhibition of rhodopsin P23H expression that is not able to prevent or block photoreceptor degeneration. Since rhodopsin is the most abundant rod photoreceptor protein, systems resulting in more robust shRNA expression in the retina may be required to achieve therapeutic efficacy in vivo.
引用
收藏
页码:692 / 699
页数:8
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