文献类型：期刊文献

英文题名：Prime editing enables precise genome modification of a Populus hybrid

作者：Zou, Jinpeng[1,2] Li, Yuhong[1] Wang, Kejian[2,3] Wang, Chun[2] Zhuo, Renying[1]

第一作者：Zou, Jinpeng

通信作者：Zhuo, RY[1];Wang, C[2]

机构：[1]Chinese Acad Forestry, Res Inst Subtrop Forestry, State Key Lab Tree Genet & Breeding, Key Lab Tree Breeding Zhejiang Prov, Hangzhou 311400, Peoples R China;[2]Chinese Acad Agr Sci, China Natl Rice Res Inst, State Key Lab Rice Biol & Breeding, Hangzhou 310006, Peoples R China;[3]Minist Agr & Rural Affairs, Key Lab Gene Editing Technol Hainan, Sanya 572025, Peoples R China

年份：2024

卷号：5

期号：4

起止页码：497-501

外文期刊名：ABIOTECH

收录：EI(收录号：20243717016703);Scopus(收录号：2-s2.0-85203279036);WOS:【ESCI(收录号:WOS:001308073400001)】；

基金：We thank Professor Mengzhu Lu from Zhejiang A&F University for providing the 84K callus. This work was 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) and the Central Public-interest Scientific Institution Basal Research Fund, China (CPSIBRF-CNRRI-202203).

语种：英文

外文关键词：Poplar; Prime editing; PE3 system; Dicot

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 x P. glandulosa). We used the 2 x 35S 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 T-0 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.