引入 PRPH2 c.828+1G>A 突变的 Prime 编辑策略。

Prime Editing Strategy to Install the PRPH2 c.828+1G>A Mutation.

机构信息

Department of Biomedical Engineering, Columbia University, New York, NY, USA.

Edward S. Harkness Eye Institute, Department of Ophthalmology, Columbia University Irving Medical Center/New York-Presbyterian Hospital, New York, NY, USA.

出版信息

Adv Exp Med Biol. 2023;1415:97-102. doi: 10.1007/978-3-031-27681-1_15.

DOI:10.1007/978-3-031-27681-1_15

PMID:37440020

Abstract

Mutations in peripherin 2 (PRPH2) are associated with a spectrum of inherited retinal diseases (IRDs) including retinitis pigmentosa (RP) and macular degeneration. As PRPH2 is localized to cone and rod outer segments, mutations in PRPH2 lead the disorganization or absence of photoreceptor outer segments. Here, we report on a patient with PRPH2-linked RP who exhibited widespread RPE atrophy with a central area of macular atrophy sparing the fovea. In future studies, we plan to model the pathobiology of PRPH2-based RP using induced pluripotent stem cell (iPSC)-derived retinal organoids. To effectively model rare mutations using iPSC-derived retinal organoids, we first require a strategy that can install the desired mutation in healthy wild-type iPSC, which can efficiently generate well-laminated retinal organoids. In this study, we developed an efficient prime editing strategy for the installation of the pathogenic PRPH2 c.828+1 G>A splice-site mutation underlying our patient's disease.

摘要

外周蛋白 2（PRPH2）突变与一系列遗传性视网膜疾病（IRDs）相关，包括色素性视网膜炎（RP）和黄斑变性。由于 PRPH2 定位于视锥和视杆外节，PRPH2 突变导致光感受器外节的紊乱或缺失。在这里，我们报告了一位 PRPH2 相关 RP 患者，其表现为广泛的 RPE 萎缩，伴有黄斑中心区萎缩，避开了中央凹。在未来的研究中，我们计划使用诱导多能干细胞（iPSC）衍生的视网膜类器官来模拟基于 PRPH2 的 RP 的病理生物学。为了使用 iPSC 衍生的视网膜类器官有效地模拟罕见突变，我们首先需要一种能够在健康的野生型 iPSC 中安装所需突变的策略，该策略能够有效地生成分层良好的视网膜类器官。在这项研究中，我们开发了一种有效的 Prime 编辑策略，用于安装我们患者疾病相关的致病性 PRPH2 c.828+1 G>A 剪接位点突变。