CRISPR/Cas9-Mediated Deletion of Large Genomic Fragments in Soybean

文献类型： 外文期刊

第一作者： Cai, Yupeng

作者： Cai, Yupeng;Chen, Li;Yao, Weiwei;Hou, Wensheng;Cai, Yupeng;Chen, Li;Sun, Shi;Wu, Cunxiang;Yao, Weiwei;Jiang, Bingjun;Han, Tianfu;Hou, Wensheng

作者机构：

关键词： Agrobacterium tumefaciens-mediated transformation; CRISPR/Cas9; dual-sgRNA/Cas9 design; large fragment deletion; soybean

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2018 年 19 卷 12 期

页码：

收录情况： SCI

摘要： At present, the application of CRISPR/Cas9 in soybean (Glycine max (L.) Merr.) has been mainly focused on knocking out target genes, and most site-directed mutagenesis has occurred at single cleavage sites and resulted in short deletions and/or insertions. However, the use of multiple guide RNAs for complex genome editing, especially the deletion of large DNA fragments in soybean, has not been systematically explored. In this study, we employed CRISPR/Cas9 technology to specifically induce targeted deletions of DNA fragments in GmFT2a (Glyma16g26660) and GmFT5a (Glyma16g04830) in soybean using a dual-sgRNA/Cas9 design. We achieved a deletion frequency of 15.6% for target fragments ranging from 599 to 1618 bp in GmFT2a. We also achieved deletion frequencies of 12.1% for target fragments exceeding 4.5 kb in GmFT2a and 15.8% for target fragments ranging from 1069 to 1161 bp in GmFT5a. In addition, we demonstrated that these CRISPR/Cas9-induced large fragment deletions can be inherited. The T2 transgene-free' homozygous ft2a mutants with a 1618 bp deletion exhibited the late-flowering phenotype. In this study, we developed an efficient system for deleting large fragments in soybean using CRISPR/Cas9; this system could benefit future research on gene function and improve agriculture via chromosome engineering or customized genetic breeding in soybean.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序