Targeted mutagenesis in soybean using the CRISPR-Cas9 system

doi:10.1038/srep10342

	Targeted mutagenesis in soybean using the CRISPR-Cas9 system
	Sun, Xianjun 1,2; Hu, Zheng 2; Chen, Rui 3; Jiang, Qiyang 2; Song, Guohua 2; Zhang, Hui 2; Xi, Yajun 1
刊名	SCIENTIFIC REPORTS
	2015
卷号	5 页码:-
ISSN号	2045-2322
DOI	10.1038/srep10342
通讯作者	Sun, Xianjun
英文摘要	Genome editing is a valuable technique for gene function analysis and crop improvement. Over the past two years, the CRISPR-Cas9 system has emerged as a powerful tool for precisely targeted gene editing. In this study, we predicted 11 U6 genes in soybean (Glycine max L.). We then constructed two vectors (pCas9-GmU6-sgRNA and pCas9-AtU6-sgRNA) using the soybean U6-10 and Arabidopsis U6-26 promoters, respectively, to produce synthetic guide RNAs (sgRNAs) for targeted gene mutagenesis. Three genes, Glyma06g14180, Glyma08g02290 and Glyma12g37050, were selected as targets. Mutations of these three genes were detected in soybean protoplasts. The vectors were then transformed into soybean hairy roots by Agrobacterium rhizogenes infection, resulting in efficient target gene editing. Mutation efficiencies ranged from 3.2-9.7% using the pCas9-AtU6-sgRNA vector and 14.7-20.2% with the pCas9-GmU6-sgRNA vector. Biallelic mutations in Glyma06g14180 and Glyma08g02290 were detected in transgenic hairy roots. Off-target activities associated with Glyma06g14180 and Glyma12g37050 were also detected. Off-target activity would improve mutation efficiency for the construction of a saturated gene mutation library in soybean. Targeted mutagenesis using the CRISPR-Cas9 system should advance soybean functional genomic research, especially that of genes involved in the roots and nodules.
学科主题	Multidisciplinary Sciences
语种	英语
出版者	NATURE PUBLISHING GROUP
WOS记录号	WOS:000355587600001
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/4710]
专题	作物科学研究所_种质资源保存与研究中心
作者单位	1.Northwest A&F Univ, Coll Agron, Yangling 712100, Shaanxi, Peoples R China 2.Chinese Acad Agr Sci, Inst Crop Sci, Natl Key Facil Crop Genet Resources & Improvement, Beijing 100081, Peoples R China 3.Tianjin Acad Agr Sci, Tianjin Inst Agr Qual Stand & Testing Technol, Tianjin 300381, Peoples R China
推荐引用方式 GB/T 7714	Sun, Xianjun,Hu, Zheng,Chen, Rui,et al. Targeted mutagenesis in soybean using the CRISPR-Cas9 system[J]. SCIENTIFIC REPORTS,2015,5:-.
APA	Sun, Xianjun.,Hu, Zheng.,Chen, Rui.,Jiang, Qiyang.,Song, Guohua.,...&Xi, Yajun.(2015).Targeted mutagenesis in soybean using the CRISPR-Cas9 system.SCIENTIFIC REPORTS,5,-.
MLA	Sun, Xianjun,et al."Targeted mutagenesis in soybean using the CRISPR-Cas9 system".SCIENTIFIC REPORTS 5(2015):-.