基于双碱基编辑系统的植物基因靶向随机突变技术  

Saturation Mutagenesis Using Dual Cytosine and Adenine Base Editors

作　　者：张瑞[1] 高彩霞[1] Rui Zhang;Caixia Gao(Center for Genome Editing,State Key Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics and Developmental Biology,Innovation Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China)

机构地区：[1]中国科学院种子创新研究院,遗传与发育生物学研究所,植物细胞与染色体工程国家重点实验室,基因组编辑中心,北京100101

出　　处：《植物学报》2021年第1期50-55,共6页Chinese Bulletin of Botany

基　　金：中国科学院战略性先导专项(No.XDA24020102);国家自然科学基金(No.31788103,No.31900301);国家重点研发计划(No.2016 YFD0101804)。

摘　　要：遗传性变异是表型多样性的基础,靶向饱和突变作物基因可以促进产生具有优异农艺性状的突变体。相较于传统诱变育种和异源物种中的定向进化方法,基于双碱基编辑系统的植物基因靶向随机突变技术可对植物内源基因产生高效突变,从而实现原位定向进化,加快植物育种及功能基因研究进程。该文介绍了使用饱和靶向内源基因突变编辑器(STEME)对植物功能基因进行靶向随机突变的具体操作方法,从靶点设计和突变体检测方面进行了详细描述。Because the genome of an organism determines its primary phenotype,evolutionary principles suggest that genetic variations enhance phenotypic diversity towards increased fitness.Targeted saturation mutagenesis of crop genes could be used to screen for genetic variants with improved agronomic traits.Compared to traditional mutational breeding or directed evolution in heterologous organisms,targeted mutagenesis via dual cytosine and adenine base editors effectively generates endogenous mutagenesis and facilitates in vivo directed evolution of plant genes.In this protocol,we detail the process towards using saturated targeted endogenous mutagenesis editors(STEMEs)to generate targeted,random mutagenesis of plant genes.In particular,we focus on the process of designing targets,screening and genotyping the resulting evolved variants.

关 键 词：基因编辑 碱基编辑 靶向随机突变 植物基因定向进化 

分 类 号：Q943.2[生物学—植物学]