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

文献类型： 外文期刊

第一作者： Zhang, Wenxin

作者： Zhang, Wenxin;Wang, Rui;Kong, Dali;Peng, Fangnan;Chen, Mei;Zeng, Wenjie;He, Sheng;Wang, Zhen;Zhu, Jian-Kang;Miki, Daisuke;Zhang, Wenxin;Wang, Rui;Kong, Dali;Peng, Fangnan;Zeng, Wenjie;Giaume, Francesca;Fornara, Fabio;Zhang, Hui;Kyozuka, Junko;Zhu, Jian-Kang;Zhu, Jian-Kang;Zhu, Jian-Kang;Zhang, Wenxin;Wang, Zhen

作者机构：

期刊名称：CELL REPORTS METHODS （ 影响因子：3.8； 五年影响因子：3.8 ）

ISSN： 2667-2375

年卷期： 2023 年 3 卷 1 期

页码：

收录情况： SCI

摘要： 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.

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