Preferential silencing of a common dominant rhodopsin mutation does not inhibit retinal degeneration in a transgenic model.
Preferential silencing of a common dominant rhodopsin mutation does not inhibit retinal degeneration in a transgenic model.
复制标题
在转基因模型中，常见显性视紫红质突变的优先沉默不会抑制视网膜变性。
DOI：
10.1016/j.ymthe.2006.07.008
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Auricchio,Alberto
中科院分区：
文献类型：
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作者：
Tessitore,Alessandra;Parisi,Fabiana;Denti,MichelaAlessandra;Allocca,Mariacarmela;DiVicino,Umberto;Domenici,Luciano;Bozzoni,Irene;Auricchio,Alberto
常染色体显性视网膜色素变性由频繁的视紫红质P23H突变引起，其特征是进行性光感受器细胞死亡，最终导致失明，目前尚无治疗方法。考虑到P23H突变所产生的功能获得效应，旨在沉默突变等位基因表达的策略，如RNA干扰，是可取的。我们设计了小干扰rna （siRNA）来特异性沉默P23H视紫红质等位基因，该等位基因是由转基因大鼠疾病模型表达的。我们选择了干扰素siRNA，并基于所选择的siRNA生成了表达短发夹RNA （shRNA）的腺相关病毒（AAV）载体。在体外，shRNA显著抑制P23H而非野生型视紫红质等位基因的表达。在P23H转基因大鼠的视网膜下给予编码shRNA的AAV2/5载体可抑制视紫红质P23H的表达，但不能阻止或阻止光感受器变性。由于视紫红质是最丰富的视杆光感受器蛋白，因此可能需要在视网膜中产生更强shRNA表达的系统来实现体内治疗效果。
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 selectedin vitroone siRNA and generated an adeno-associated viral (AAV) vector expressing the short hairpin RNA (shRNA) based on the selected siRNA.In vitrothe 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 efficacyin vivo.