In Vivo CRISPR/Cas9-Mediated Genome Editing Mitigates Photoreceptor Degeneration in a Mouse Model of X-Linked Retinitis Pigmentosa
In Vivo CRISPR/Cas9-Mediated Genome Editing Mitigates Photoreceptor Degeneration in a Mouse Model of X-Linked Retinitis Pigmentosa
复制标题
体内 CRISPR/Cas9 介导的基因组编辑减轻 X 连锁视网膜色素变性小鼠模型中的光感受器变性
DOI：
10.1167/iovs.61.4.31
复制
发表时间：
2020-04-01
影响因子：
4.4
通讯作者：
Yang, Liping
中科院分区：
文献类型：
--
作者：
Hu, Shuang;Du, Juan;Yang, Liping
目的。色素性视网膜炎 GTP 酶调节因子 (RPGR) 相关的 X 连锁色素性视网膜炎与遗传性视网膜疾病中最严重的表型之一相关。本研究的目的是在 Rpgr.METHODS 小鼠模型中研究成簇规则间隔短回文重复/Cas9 介导的基因编辑疗法。 Rpgr(-/y)Cas9(+/WT) 雄性小鼠用于本研究。 6月龄时，分别接受单次视网膜下注射携带sgRNA的腺相关病毒载体和供体模板，并在1、6和12个月后检查治疗效果。结果。 Rpgr 基因敲除小鼠表现出缓慢但渐进的与年龄相关的视网膜变性，这与人类发生的疾病相似。在 6 个月和 12 个月后，在治疗的视网膜部分观察到显着的光感受器保留，与同一只眼睛的未治疗的视网膜部分形成鲜明对比。令人惊讶的是，在有丝分裂后光感受器中观察到了基因组 DNA 测序所证明的目标基因座的精确修饰。而且，治疗效果可持续长达12个月，且未出现脱靶效应。结论。我们的研究有力地表明，基因编辑疗法是治疗遗传性视网膜变性的一种有前途的治疗策略。
PURPOSE. Retinitis pigmentosa GTPase regulator (RPGR)-related X-linked retinitis pigmentosa is associated with one of the most severe phenotypes among inherited retinal disease. The aim of this study was to investigate Clustered Regularly Interspaced Short Palindromic Repeat/Cas9-mediated gene editing therapy in a mouse model of Rpgr.METHODS. The Rpgr(-/y)Cas9(+/WT) male mice were used for this study. At 6 months of age, they received a single subretinal injection of adeno-associated virus vectors carrying sgRNA and donor template separately, and therapeutic effect was examined after 1, 6, and 12 months.RESULTS. Rpgr knockout mouse showed slow but progressive age-related retinal degeneration, which emulates the disease occurring in humans. Significant photoreceptor preservation was observed in the treated part of the retina, in sharp contrast to the untreated part of the retina in the same eye after 6 and 12 months. It was surprising that precise modification at the target locus as demonstrated by genomic DNA sequencing in the post-mitotic photoreceptor was observed. Moreover, the therapeutic effect lasts for up to 12 months and no off-target effects were shown.CONCLUSIONS. Our study strongly demonstrates that gene editing therapy is a promising therapeutic strategy to treat inherited retinal degeneration.