基于双碱基编辑系统的植物基因靶向随机突变技术
收稿日期: 2021-01-11
录用日期: 2021-02-25
网络出版日期: 2021-02-25
基金资助
中国科学院战略性先导专项(XDA24020102);国家自然科学基金(31788103);国家自然科学基金(31900301);国家重点研发计划(2016 YFD0101804)
Saturation Mutagenesis Using Dual Cytosine and Adenine Base Editors
Received date: 2021-01-11
Accepted date: 2021-02-25
Online published: 2021-02-25
张瑞, 高彩霞 . 基于双碱基编辑系统的植物基因靶向随机突变技术[J]. 植物学报, 2021 , 56(1) : 50 -55 . DOI: 10.11983/CBB21009
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.
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