摘要
人类众多遗传性疾病是由于基因组突变所导致,而基于基因编辑技术的基因治疗策略,可从DNA层面对致病突变进行彻底修复,为疾病的治愈提供了新的可能.随着基因编辑技术的不断发展与突破,2019年新型DNA编辑工具先导编辑(PE)成功问世.PE技术并不会引起DNA双链断裂(DSBs),也无需单独引入DNA模板,即可实现基因编辑,可广泛应用于包括点突变及小片段的插入、删除等不同场景.与传统基因编辑工具相比,PE因其高编辑效率及低脱靶率,具备更高的有效性与安全性.本篇综述:(1)详细介绍了先导编辑的工作原理及发展历史.(2)概述了先导编辑系统的改良策略,包括相关酶的改造、工具RNA的结构改进以及递送方式的创新(3)梳理了辅助PE系统设计和分析的网页工具,简化了PE系统的搭建及使用流程.(4)系统总结了PE在疾病模型中的应用案例,并展望了未来的临床应用前景.
Gene editing ushers in a new era of disease treatment since many genetic diseases are caused by basepair mutations in genomic DNA.With the rapid development of genome editing technology,novel editing tools such as base editing and prime editing(PE)have attracted public attention,heralding a great leap forward in this field.PE,in particular,is characterized by no need for double-strand breaks(DSBs)or homology sequence templates with variable application scenarios,including point mutations as well as insertions or deletions.With higher editing efficiency and fewer byproducts than traditional editing tools,PE holds great promise as a therapeutic strategy for human diseases.Subsequently,a growing demand for the standard construction of PE system has spawned numerous easy-to-access internet resources and tools for personalized prime editing guide RNA(pegRNA)design and off-target site prediction.In this review,we mainly introduce the innovation and evolutionary strategy of PE systems and the auxiliary tools for PE design and analysis.Additionally,its application and future potential in the clinical field have been summarized and envisaged.
作者
傅伊甸
何晓雨
高鑫
李芳
葛盛芳
阳志
范先群
Yidian Fu;Xiaoyu He;Xin D.Gao;Fang Li;Shengfang Ge;Zhi Yang;Xianqun Fan(Department of Ophthalmology,Ninth People’s Hospital,Shanghai JiaoTong University School of Medicine,Shanghai 200011,China;Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology,Shanghai 200011,China;Merkin Institute of Transformative Technologies in Healthcare,Broad Institute of Harvard and MIT,Cambridge MA 02141,USA;Department of Chemistry and Chemical Biology,Harvard University,Cambridge MA 02138,USA;Howard Hughes Medical Institute,Harvard University,Cambridge MA 02138,USA)
基金
supported by the National Basic Science Center Program of China(82388101)
the National Natural Science Foundation of China(82200961 and 82203260)
the Science and Technology Commission of Shanghai(20DZ2270800)
Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology(2022SKLEKFKT004)
China Postdoctoral Science Foundation(2022M720091)。
