AAV 递送的 CRISPR/Cas9 基因敲除可预防小鼠视网膜变性。

Nrl knockdown by AAV-delivered CRISPR/Cas9 prevents retinal degeneration in mice.

机构信息

Ocular Gene Therapy Core, National Eye Institute, NIH, 6 Center Drive, Room 307, Bethesda, Maryland 20892, USA.

Neurobiology-Neurodegeneration and Repair Laboratory, National Eye Institute, NIH, 6 Center Drive, Room 307, Bethesda, Maryland 20892, USA.

出版信息

Nat Commun. 2017 Mar 14;8:14716. doi: 10.1038/ncomms14716.

DOI:10.1038/ncomms14716

PMID:28291770

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5355895/

Abstract

In retinitis pigmentosa, loss of cone photoreceptors leads to blindness, and preservation of cone function is a major therapeutic goal. However, cone loss is thought to occur as a secondary event resulting from degeneration of rod photoreceptors. Here we report a genome editing approach in which adeno-associated virus (AAV)-mediated CRISPR/Cas9 delivery to postmitotic photoreceptors is used to target the Nrl gene, encoding for Neural retina-specific leucine zipper protein, a rod fate determinant during photoreceptor development. Following Nrl disruption, rods gain partial features of cones and present with improved survival in the presence of mutations in rod-specific genes, consequently preventing secondary cone degeneration. In three different mouse models of retinal degeneration, the treatment substantially improves rod survival and preserves cone function. Our data suggest that CRISPR/Cas9-mediated NRL disruption in rods may be a promising treatment option for patients with retinitis pigmentosa.

摘要

在色素性视网膜炎中，视锥细胞感光器的丧失会导致失明，而保留视锥细胞功能是一个主要的治疗目标。然而，视锥细胞的丧失被认为是继发于杆状细胞感光器变性的事件。在这里，我们报告了一种基因组编辑方法，即通过腺相关病毒（AAV）介导的 CRISPR/Cas9 传递到有丝分裂后感光器，用于靶向 Nrl 基因，该基因编码神经视网膜特异性亮氨酸拉链蛋白，是感光器发育过程中杆状细胞命运决定因素。在 Nrl 被破坏后，杆状细胞获得了部分视锥细胞的特征，并在存在杆状细胞特异性基因突变的情况下表现出更好的存活率，从而防止了继发的视锥细胞变性。在三种不同的视网膜变性小鼠模型中，该治疗方法显著提高了杆状细胞的存活率并保留了视锥细胞的功能。我们的数据表明，CRISPR/Cas9 介导的 NRL 破坏在杆状细胞中可能是治疗色素性视网膜炎患者的一种有前途的治疗选择。