Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep

doi:10.1038/srep32271

	Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep
	Wang, Xiaolong 1; Niu, Yiyuan 1; Zhou, Jiankui2,3 ; Yu, Honghao 4; Kou, Qifang 5; Lei, Anmin 6; Zhao, Xiaoe 6; Yan, Hailong 1,4; Cai, Bei 1; Shen, Qiaoyan 6; Zhou, Shiwei 1; Zhu, Haijing 4; Zhou, Guangxian 1; Niu, Wenzhi 5; Hua, Jinlian 6; Jiang, Yu 1; Huang, Xingxu2,3 ; Ma, Baohua 6; Chen, Yulin 1
	2016-08-26
发表期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
卷号	6
发表状态	已发表
DOI	10.1038/srep32271
摘要	The CRISPR/Cas9 system provides a flexible approach for genome engineering of genetic loci. Here, we successfully achieved precise gene targeting in sheep by co-injecting one-cell-stage embryos with Cas9 mRNA and RNA guides targeting three genes (MSTN, ASIP, and BCO2). We carefully examined the sgRNAs: Cas9-mediated targeting effects in injected embryos, somatic tissues, as well as gonads via cloning and sequencing. The targeting efficiencies in these three genes were within the range of 27-33% in generated lambs, and that of simultaneously targeting the three genes was 5.6%, which demonstrated that micro-injection of zygotes is an efficient approach for generating gene-modified sheep. Interestingly, we observed that disruption of the MSTN gene resulted in the desired muscle hypertrophy that is characterized by enlarged myofibers, thereby providing the first detailed evidence supporting that gene modifications had occurred at both the genetic and morphological levels. In addition, prescreening for the off-target effect of sgRNAs was performed on fibroblasts before microinjection, to ensure that no detectable off-target mutations from founder animals existed. Our findings suggested that the CRISPR/Cas9 method can be exploited as a powerful tool for livestock improvement by simultaneously targeting multiple genes that are responsible for economically significant traits.
收录类别	SCI
语种	英语
资助项目	China Agriculture Research System[CARS-40-13]
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000381971700001
出版者	NATURE PUBLISHING GROUP
WOS关键词	ONE-STEP GENERATION ; SIGNALING PROTEIN ASIP ; MUTATION ; EXPRESSION ; WHITE ; MICE ; PLANTS
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1747
专题	生命科学与技术学院_PI研究组_黄行许组
通讯作者	Huang, Xingxu; Ma, Baohua; Chen, Yulin
作者单位	1.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Peoples R China 2.Nanjing Univ, Natl Resource Ctr Mutant Mice, MOE Key Lab Model Anim Dis Study, Model Anim Res Ctr, Nanjing 210061, Jiangsu, Peoples R China 3.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 4.Yulin Univ, Coll Life Sci, Yulin 719000, Peoples R China 5.Ningxia Tianyuan Sheep Farm, Hongsibu 751999, Peoples R China 6.Northwest A&F Univ, Coll Vet Med, Yangling 712100, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Wang, Xiaolong,Niu, Yiyuan,Zhou, Jiankui,et al. Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep[J]. SCIENTIFIC REPORTS,2016,6.
APA	Wang, Xiaolong.,Niu, Yiyuan.,Zhou, Jiankui.,Yu, Honghao.,Kou, Qifang.,...&Chen, Yulin.(2016).Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep.SCIENTIFIC REPORTS,6.
MLA	Wang, Xiaolong,et al."Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep".SCIENTIFIC REPORTS 6(2016).