Intron-targeted gene insertion in rice using CRISPR/Cas9: A case study of the Pi-ta gene

文献类型： 外文期刊

第一作者： Xu, Yang

作者： Xu, Yang;Wang, Fangquan;Chen, Zhihui;Wang, Jun;Li, Wen-Qi;Fan, Fangjun;Tao, Yajun;Zhao, Ling;Zhong, Weigong;Yang, Jie;Xu, Yang;Wang, Fangquan;Chen, Zhihui;Wang, Jun;Li, Wen-Qi;Fan, Fangjun;Zhong, Weigong;Yang, Jie;Xu, Yang;Wang, Fangquan;Yang, Jie;Zhu, Qian-Hao;Yang, Jie

作者机构：

关键词： Rice; CRISPR/Cas9; Pi-ta; Genome editing; Molecular breeding

期刊名称：CROP JOURNAL （ 影响因子：4.407； 五年影响因子：5.687 ）

ISSN： 2095-5421

年卷期： 2020 年 8 卷 3 期

页码：

收录情况： SCI

摘要： Intron-targeted gene insertion strategy using CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated Cas9) has been shown to be a potential tool for crop genetic improvement by targeted mutagenesis or gene replacement of an elite allele into widely cultivated rice varieties. The rice blast resistant protein Pi-ta, differs from its susceptible counterpart, pi-ta, by a single amino acid in exon 2. To create new materials resistant to the rice blast disease, we inserted a genomic fragment containing the exon 2 and 3' untranslated region (3' UTR) of Pi-ta into intron 1 of pi-ta in rice materials susceptible to rice blast using the intron-targeted insertion strategy. The gene insertion frequency was 3.8%. Several novel transgene-free progeny with stably inherited homozygous insert were identified in the T-1 generation, which have been crossed to rice germplasm bearing other resistance gene (R gene) for pyramiding of R genes. This work verified the feasibility of using the genome editing technology in improvement of qualitative agronomic trait in crops. (C) 2019 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

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