Dual-AAV split prime editor corrects the mutation and phenotype in mice with inherited retinal degeneration  

作　　者：Kaiqin She Yi Liu Qinyu Zhao Xiu Jin Yiliu Yang Jing Su Ruiting Li Li Song Jianlu Xiao Shaohua Yao Fang Lu Yuquan Wei Yang Yang 

机构地区：[1]Department of Ophthalmology,West China Hospital,Sichuan University,Chengdu,Sichuan,China [2]State Key Laboratory of Biotherapy and Cancer Center,West China Hospital,Sichuan University and Collaborative Innovation Center,Chengdu,Sichuan,China

出　　处：《Signal Transduction and Targeted Therapy》2023年第3期1186-1197,共12页信号转导与靶向治疗（英文）

基　　金：This work was supported by the National Natural Science Foundation of China(nos.U19A2002,82222030 and 82201212).

摘　　要：The prime editor(PE)can edit genomes with almost any intended changes,including all 12 possible types of base substitutions,small insertions and deletions,and their combinations,without the requirement for double strand breaks or exogenous donor templates.PE demonstrates the possibility of correcting a variety of disease-causing mutations and might expand the therapeutic application of gene editing.In this study,PE was optimized based on a dual-adeno-associated virus(AAV)split-intein system in vitro by screening different split sites and split inteins.We found that splitting PE before amino acid 1105(Ser)of SpCas9 with Rma intein resulted in the highest on-target editing.The orientations of pegRNA and nicking sgRNA in the AAV vector were further optimized.To test the in vivo performance of the optimized dual-AAV split-PE3,it was delivered by subretinal injection in rd12 mice with inherited retinal disease Leber congenital amaurosis.The prime editors corrected the pathogenic mutation with up to 16%efficiency in a precise way,with no detectable off-target edits,restored RPE65 expression,rescued retinal and visual function,and preserved photoceptors.Our findings establish a framework for the preclinical development of PE and motivate further testing of PE for the treatment of inherited retinal diseases caused by various mutations.

关 键 词：EDITOR DEGENERATION precise 

分 类 号：R774.1[医药卫生—眼科]