Directed evolution can be rapidly applied for engineering proteins, studying gene functions, and obtaining mutants with important agronomic traits. Recently, Caixia Gao and Jiayang Li's team from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, worked together to engineer novel saturated targeted endogenous mutagenesis editors (STEMEs), realizing in vivo directed evolution and function selection in plants. This system integrated the existing two single-base editing techniques, successfully induced C:G>T:A and A:T>G:C double-base editing in plants, and artificially evolved into herbicide-resistant rice through targeting the OsACC carboxyltransferase domain coding sequence. This new method of gene directed evolution in vivo displays great application potential in important agronomic trait screening and plant functional gene researches. Here we introduce the composition, editing efficiency, and application principle of the STEME system, and compare it with the existing directed evolution methods, so as to provide a reference for accelerating the innovation of crop germplasm resources.
通过定向进化(directed evolution)可以快速进行蛋白工程改良及重要基因功能研究,以获得新型农艺性状突变体。近期,中国科学院遗传与发育生物学研究所高彩霞团队和李家洋团队合作构建了新型的饱和靶向内源诱变编辑器(saturated targeted endogenous mutagenesis editors, STEMEs),并在植物中实现了基因的定向进化和功能筛选。该系统融合了现有的2种单碱基编辑技术,成功实现在植物体内同时诱导C:G>T:A、A:T>G:C双碱基编辑,通过靶向OsACC羧基转移酶结构域编码序列定向进化出水稻除草剂抗性植株。这种在体内进行基因定向进化的新方法,对于今后农作物重要农艺性状的筛选和功能基因研究具有重要作用。本文对STEME系统的组成、编辑效率和应用原理进行介绍,并与已有的定向进化方法进行比较,为加速作物种质资源创新研究提供参考。.
Keywords: STEME; directed evolution; double base editing; germplasm innovation.