AAV-mediated knockdown of Peripherin-2 in vivo using miRNA-based hairpins
AAV-mediated knockdown of Peripherin-2 in vivo using miRNA-based hairpins
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DOI：
10.1038/gt.2009.162
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发表时间：
2010-04-01
期刊：
影响因子：
5.1
通讯作者：
Ali, R. R.
中科院分区：
文献类型：
--
作者：
Georgiadis, A.;Tschernutter, M.;Ali, R. R.
遗传性视网膜变性的基因治疗，即突变等位基因的表达对光感受器细胞具有功能获得效应，很可能依赖于突变等位基因的有效沉默。外周蛋白-2(Peripherin-2，Prph2，又称外周蛋白/RDS)是一个大量表达的光感受器特异性基因。在人类中，PRPH2的功能获得突变导致常染色体显性遗传性视网膜色素变性和显性黄斑病变。针对这些情况的基因沉默策略包括短发夹状RNA(ShRNAs)的RNA干扰。最近的证据表明，基于microRNA(MiRNA)的发夹可能提供一种更安全、更有效的替代方案。在这项研究中，我们首次使用病毒转移的基于miRNA的发夹来沉默小鼠视网膜中的Prph2。结果表明，在AAV2/8介导的视网膜下注射后3周，基于miRNA的shRNA可以在体内有效且特异性地沉默Prph2，导致5周后光感受器细胞减少近50%。我们的结论是，基于miRNA的发夹可以在有效的载体介导的视网膜递送后实现快速而稳健的基因沉默。使用基于miRNA的模板来提高发夹的沉默效率的原理可能被证明对等位基因特异性沉默有价值，其中RNAi靶点的选择有限，并为显性视网膜病变的治疗提供了一种替代治疗策略。基因治疗(2010年)17486-493；DOI：10.1038/gt.2009.162；2009年12月10日在线发布
Gene therapy for inherited retinal degeneration in which expression of a mutant allele has a gain-of-function effect on photoreceptor cells is likely to depend on efficient silencing of the mutated allele. Peripherin-2 (Prph2, also known as peripherin/RDS) is an abundantly expressed photoreceptor-specific gene. In humans, gain-of-function mutations in PRPH2 result in both autosomal dominant retinitis pigmentosa and dominant maculopathies. Gene-silencing strategies for these conditions include RNA interference by short hairpin RNAs (shRNAs). Recent evidence suggests that microRNA (miRNA)-based hairpins may offer a safer and more effective alternative. In this study, we used for the first time a virally transferred miRNA-based hairpin to silence Prph2 in the murine retina. The results show that an miRNA-based shRNA can efficiently and specifically silence Prph2 in vivo as early as 3 weeks after AAV2/8-mediated subretinal delivery, leading to a nearly 50% reduction of photoreceptor cells after 5 weeks. We conclude that miRNA-based hairpins can achieve rapid and robust gene silencing after efficient vector-mediated delivery to the retina. The rationale of using an miRNA-based template to improve the silencing efficiency of a hairpin may prove valuable for allele-specific silencing in which the choice for an RNAi target is limited and offers an alternative therapeutic strategy for the treatment of dominant retinopathies. Gene Therapy (2010) 17, 486-493; doi: 10.1038/gt.2009.162; published online 10 December 2009