Precise and heritable gene targeting in rice using a sequential transformation strategy

Zhang, Wenxin1,2,9; Wang, Rui1,2; Kong, Dali1,2; Peng, Fangnan1,2; Chen, Mei1; Zeng, Wenjie1,2; Giaume, Francesca3; He, Sheng1; Zhang, Hui4; Wang, Zhen1,10; Kyozuka, Junko5; Zhu, Jian-Kang1,6,7,8; Fornara, Fabio3; Miki, Daisuke1

2023-01-23

10.1016/j.crmeth.2022.100389

CELL REPORTS METHODS   Impact Factor & Quartile

2667-2375

Gene targeting (GT) is a powerful tool for modifying endogenous genomic sequences of interest, such as sequence replacement and gene knockin. Although the efficiency of GT is extremely low in higher plants, engineered sequence-specific nucleases (SSNs)-mediated double-strand breaks (DSBs) can improve GT frequency. We recently reported a CRISPR-Cas9-mediated approach for heritable GT in Arabidopsis, called the "sequential transformation"strategy. For efficient establishment of GT via the sequential trans-formation method, strong Cas9 activity and robust DSBs are required in the plant cells being infected with Agrobacterium carrying sgRNA and donor DNA. Accordingly, we generated two independent parental lines with maize Ubiquitin 1 promoter-driven Cas9 and established sequential transformation -mediated GT in the Japonica rice cultivar Oryza sativa Nipponbare. We achieved precise GFP knockin into the endogenous OsFTL1 and OsROS1a loci. We believe that our GT technology could be widely utilized in rice research and breeding applications.

ESCI

English

Others

National Natural Science Foundation of China (NSFC)[32188102] ; Shanghai Science and Technology Innovation Plan[20ZR1467000]

Biochemistry & Molecular Biology ; Cell Biology

Biochemical Research Methods ; Cell Biology

WOS:000925840400001

ELSEVIER

Web of Science

1.Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, CAS Ctr Excellence Mol Plant Sci, Shanghai 200032, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Univ Milan, Dept Biosci, Via Celoria 26, I-20133 Milan, Italy
4.Shanghai Normal Univ, Coll Life Sci, Shanghai 200234, Peoples R China
5.Tohoku Univ, Grad Sch Life Sci, Sendai, Miyagi 9808577, Japan
6.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen 518055, Peoples R China
7.Southern Univ Sci & Technol, Sch Life Sci, Shenzhen 518055, Peoples R China
8.Chinese Acad Agr Sci, Ctr Adv Bioind Technol, Beijing 100081, Peoples R China
9.Peking Univ, Acad Adv Interdisciplinary Studies, Peking Tsinghua Ctr Life Sci, Beijing 100871, Peoples R China
10.Anhui Agr Univ, Sch Life Sci, Hefei 230036, Peoples R China

Zhang, Wenxin,Wang, Rui,Kong, Dali,et al. Precise and heritable gene targeting in rice using a sequential transformation strategy[J]. CELL REPORTS METHODS,2023,3(1).

Zhang, Wenxin.,Wang, Rui.,Kong, Dali.,Peng, Fangnan.,Chen, Mei.,...&Miki, Daisuke.(2023).Precise and heritable gene targeting in rice using a sequential transformation strategy.CELL REPORTS METHODS,3(1).

Zhang, Wenxin,et al."Precise and heritable gene targeting in rice using a sequential transformation strategy".CELL REPORTS METHODS 3.1(2023).