摘要
成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关蛋白（CRISPR associated proteins，Cas）系统是细菌和古细菌在长期演化过程中形成的一种适应性免疫防御系统，可用来对抗入侵的病毒及外源DNA。随着技术的不断进展，研究者发现CRISPR/Cas9技术可用于靶向插入、替换或敲除真核细胞基因。研究者运用CRISPR/Cas9基因组编辑技术，发现了PAX6在角膜中的过度表达可导致先天性角膜上皮损伤，进一步加深了关于KRT12突变对Meesmann角膜上皮营养不良的致病机制的研究。研究还发现PAX6可通过对GJA8基因以及αA晶状体基因的敲除来进行先天性白内障动物模型的构建，有利于未来应用于先天性白内障的鉴别和病理分析。此外，研究者应用CRISPR/Cas9基因组编辑技术进一步证实了RHOS334基因突变的RHO等位基因、P23H突变的RHO基因、Y347X突变的Pde6b基因以及突变型RP9等位基因与视网膜色素变性的相关性，为突变型KCNJ13基因、突变型CEP290基因与Leber先天性黑矇的相关性提供了证据支持。利用该技术对VEGFR2基因以及TXNIP基因进行基因编辑可为眼内新生血管疾病提供新的治疗靶点，同时该技术在增生性玻璃体视网膜病变以及视网膜母细胞瘤的致病基因研究和动物模型构建中也发挥了重要的作用。本文对CRISPR/Cas9基因组编辑工具酶的发展历程、分子特点和作用机制进行总结，并概述了当前CRISPR/Cas9技术在眼科疾病研究中的进展及应用。
The clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated protein （Cas） is an adaptive immune system against invasive viruses and exogenous DNA, which is developed by bacteria and archaer during long-term evolution.With advances in technology, researchers have found that CRISPR/Cas9 system can precisely edit the genomes of eukaryotic cells through insertion, replacement or deletion of target genes.Using CRISPR/Cas9 genome editing technology, researchers found that overexpression of paired box gene 6 （PAX6） in cornea can cause congenital corneal epithelial damage; this technology promoted the research on the pathogenic mechanism of keratin 12 （KRT12） mutation in Meesmann corneal epithelial dystrophy; it has also built congenital cataract animal models by knocking out the GJA8 gene and αA lens gene, which is beneficial to the application of the diagnosis and pathological analysis of congenital cataract.In addition, the researchers have used CRISPR/Cas9 genome editing technology to confirm the correlation of RHOS334 mutated RHO allele, P23H mutated RHO gene, Y347X mutated Pde6b gene, and mutant RP9 allele with retinitis pigmentosa.The application of this technology has provided evidence to support the association of KCNJ13 gene and mutant CEP290 gene with Leber congenital amaurosis.CRISPR/Cas9 can provide target spot of intraocular neovascular diseases the targeted therapy by editing of VEGFR2 gene and TXNIP gene, and it also playse an important role in the study of pathogenic genes and establishment of animal models for proliferative vitreoretinopathy and retinoblastoma.In this review, we introduced the evolutionary history, the molecular characteristics and the mechanism of CRISPR/Cas9, and summarized its current research advances in eye diseases.
作者
范祥雨(综述)
徐建江(审校)
Fan Xiangyu;Xu Jianjiang(Department of Eye ＆ Visual Science,Eye,Ear,Nose ＆ Throat Hospital,Fudan University,Shanghai 200031,China)
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2018年第7期553-558,共6页
Chinese Journal Of Experimental Ophthalmology
基金
国家自然科学基金面上项目（81670820、81670818）
上海市卫计委新优青计划项目（2017YQ055）
上海市人才发展基金项目（2017）
