Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA; Divison of Neuroscience, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR, USA; Casey Eye Institute, Oregon Health & Science University, Portland, OR, USA.
Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA.
Stem Cell Reports. 2021 Nov 9;16(11):2690-2702. doi: 10.1016/j.stemcr.2021.09.007. Epub 2021 Oct 14.
Cases of Leber congenital amaurosis caused by mutations in CRX (LCA7) exhibit an early form of the disease and show signs of significant photoreceptor dysfunction and eventual loss. To establish a translational in vitro model system to study gene-editing-based therapies, we generated LCA7 retinal organoids harboring a dominant disease-causing mutation in CRX. Our LCA7 retinal organoids develop signs of immature and dysfunctional photoreceptor cells, providing us with a reliable in vitro model to recapitulate LCA7. Furthermore, we performed a proof-of-concept study in which we utilize allele-specific CRISPR/Cas9-based gene editing to knock out mutant CRX and saw moderate rescue of photoreceptor phenotypes in our organoids. This work provides early evidence for an effective approach to treat LCA7, which can be applied more broadly to other dominant genetic diseases.
由 CRX（LCA7）基因突变引起的莱伯先天性黑蒙症病例表现出疾病的早期形式，并显示出明显的光感受器功能障碍和最终丧失的迹象。为了建立一个用于研究基于基因编辑的治疗方法的转化体外模型系统，我们生成了携带 CRX 显性致病突变的 LCA7 视网膜类器官。我们的 LCA7 视网膜类器官表现出不成熟和功能障碍的光感受器细胞的迹象，为我们提供了一个可靠的体外模型来重现 LCA7。此外，我们进行了一项概念验证研究，我们利用等位基因特异性 CRISPR/Cas9 基于基因编辑来敲除突变的 CRX，并在我们的类器官中观察到光感受器表型的适度恢复。这项工作为治疗 LCA7 的有效方法提供了早期证据，该方法可以更广泛地应用于其他显性遗传疾病。
