Prime editing enables precise genome modification of a Populus hybrid  

作　　者：Jinpeng Zou Yuhong Li Kejian Wang Chun Wang& Renying Zhuo 

机构地区：[1]State Key Laboratory of Tree Genetics and Breeding,Key Laboratory of Tree Breeding of Zhejiang Province,Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 311400,China [2]State Key Laboratory of Rice Biology and Breeding,China National Rice Research Institute,Chinese Academy of Agricultural Sciences,Hangzhou 310006,China [3]Key Laboratory of Gene Editing Technologies(Hainan),Ministry of Agriculture and Rural Affairs,Sanya 572025,China

出　　处：《aBIOTECH》2024年第4期497-501,共5页生物技术通报(英文版)

基　　金：supported by the National Key Research and Development Program of China(2021YFD2200201);the China National Rice Research Institute Key Research and Development Project(CNRRI-2020-01);the Central Public-interest Scientific Institution Basal Research Fund,China(CPSIBRF-CNRRI-202203).

摘　　要：CRISPR/Cas-based genome editing has been extensively employed in the breeding and genetic improvement of trees,yet precise editing remains challenging in these species.Prime editing(PE),a revolutionary technology for precise editing,allows for arbitrary base substitutions and the insertion/deletion of small fragments.In this study,we focused on the model tree poplar 84K(Populus alba 9 P.glandulosa).We used the 2935S promoter to express a fusion protein of spCas9 nickase(nCas9)and engineered Moloney murine leukemia virus(MMLV),and the Arabidopsis thaliana AtU6 promoter to express an engineered PE guide RNA(epegRNA)and Nick gRNA,pioneering the establishment of the Prime Editor 3(PE3)system in dicot poplar.Single-base substitutions,multiple-base substitutions,and small-fragment insertions/deletions were edited into three endogenous target genes.The desired edits were identified in hygromycin-resistant(transformed)calli at seven out of nine target sites,with an average editing efficiency ranging from 0.1 to 3.6%.Furthermore,stable T0 plants contained the desired edits at four out of nine targets,with editing efficiencies ranging from 3.6 to 22.2%.Establishment of the PE3 system provides a powerful tool for the precise modification of the poplar genome.

关 键 词：POPLAR Prime editing PE3 system DICOT 

分 类 号：Q78[生物学—分子生物学]