引导编辑:突破碱基编辑类型的新技术  

Prime editing:a search and replace tool with versatile base changes

作　　者：刘尧 周先辉 黄舒泓 王小龙[1] Yao Liu;Xianhui Zhou;Shuhong Huang;Xiaolong Wang(College of Animal Science and Technology/International Joint Research Center of Animal Bio-breeding,Ministry of Agriculture and Rural Affairs/Shaanxi Key Laboratory of Animal Genetic Breeding and Reproduction,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学,动物科技学院/农业农村部动物生物育种国际联合研究中心/陕西省动物遗传育种与繁殖重点实验室,杨凌712100

出　　处：《遗传》2022年第11期993-1008,共16页Hereditas（Beijing)

基　　金：国家自然科学基金项目(编号:31972526,32161143010)资助。

摘　　要：引导编辑技术(prime editing)是一种基于CRISPR/Cas系统的新型基因编辑技术。引导编辑器(prime editor,PE)的效应蛋白是逆转录酶与具有单链切割活性的nCas9(H840A)的融合蛋白,被称为PE2;其向导RNA是通过在sgRNA的3′末端增加逆转录模板和引物结合位点序列构成,被称为pegRNA。PE系统可以实现所有12种类型的碱基突变,还可以实现短的插入删除编辑,及多种编辑类型的组合。自2019年被开发以来,因其编辑类型多样化及高特异性等优势,PE已经被成功地应用在多种动植物及细菌中,同时在基因治疗和农业育种等领域也展现出良好的应用前景。本文对PE系统的开发过程、特点、优化、应用、安全风险等方面进行了系统介绍,并对其发展前景进行了展望,以期有助于相关领域研究人员对PE系统的了解与应用。Prime editing is a newly developed CRISPR/Cas system-based genome editing technique.The effector of prime editor(PE)is termed PE2,which is generated by fusing a reverse transcriptase(RT)with a Cas9 H840A nickase.The guide RNA of PE is termed prime editing guide RNA(pegRNA),which consists of a single guide RNA(sgRNA)with a 3′extension containing the RT template(RTT)and primer binding site(PBS).PE can install all 12 types of point mutations,small insertions and deletions and combinations thereof.Since its emergence in 2019,with the high versatility and specificity,PE has been applied to many living organisms,including animals,plants and bacteria.This led to many explorations of PE on gene therapy and genetic improvement in agriculture.In this review,we systematically describe the development,characteristics,optimizations,applications and security of PE.In addition,we discuss the future applications of PE.We expect that this review will help researchers to grasp and better use PE.

关 键 词：引导编辑器 CRISPR/Cas9 优化 应用 

分 类 号：Q78[生物学—分子生物学]