基于CRISPR/Cas9技术的哺乳动物细胞基因敲入的策略优化与效率提升  

Strategy optimization and efficiency improvement of gene knock-in in mammalian cells based on CRISPR/Cas9 technology

作　　者：邹爽 孟德豪 李贺年 张艳芬[2] 马宁 ZOU Shuang;MENG Dehao;LI Henian;ZHANG Yanfen;MA Ning(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Harbin Medical University,Harbin 150081,China;Department of Clinical Laboratory,the Second Affiliated Hospital,Harbin Medical University,Harbin 150000,China;Key Laboratory of Genetically Modified Model Animals of Heilongjiang Province,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学基础医学院生物化学与分子生物学教研室,哈尔滨150081 [2]哈尔滨医科大学附属第二医院检验科,哈尔滨150000 [3]黑龙江省遗传改变模式动物重点实验室,哈尔滨150081

出　　处：《生命的化学》2024年第8期1433-1441,共9页Chemistry of Life

基　　金：黑龙江省省级领军人才梯队后备带头人项目(2018);哈尔滨医科大学少帅揭榜项目(HMUMIF-21025);黑龙江省教育厅教改课题重点委托项目(SJGZ20200109);哈尔滨医科大学附属第二医院中青年创新科学研究基金项目(2020)。

摘　　要：CRISPR基因编辑系统自问世以来,极大推动了疾病治疗、药物研发、疾病模型构建等领域的研究进展。基于该系统的基因敲入(knock-in,KI)策略已成为基因工程生物制备与基因治疗等应用的常用策略。但外源DNA的敲入,尤其是较大DNA片段的敲入效率一直是该策略有待优化的问题。研究者已从基因(组)编辑系统、DNA损伤修复、细胞周期调节等方面进行了提升KI效率的探索。本文主要针对以上方面进行综述,阐述影响KI效率的关键因素与提升KI效率的优化策略。Since the advent of CRISPR gene editing system,it has greatly promoted the research progress in the field of disease treatment,drug discovery,and disease model construction.The gene knock-in(KI)strategy based on this system has become a common strategy for the preparation of genetically engineered organisms and gene therapy applications.However,the tapping efficiency of exogenous DNA,especially for larger DNA fragments,has always been a problem that needs to be optimized in this strategy.Researchers have explored how to improve KI efficiency from the aspects of gene(group)editing system,DNA damage repair,cell cycle regulation.This paper mainly reviews the above aspects,describes the key factors affecting KI efficiency and optimization strategies for improving KI efficiency.

关 键 词：CRISPR/Cas9 基因敲入 同源重组修复 非同源末端连接 

分 类 号：Q78[生物学—分子生物学]