Trio-Based Deep Sequencing Reveals a Low Incidence of Off-Target Mutations in the Offspring of Genetically Edited Goats

doi:10.3389/fgene.2018.00449

	Trio-Based Deep Sequencing Reveals a Low Incidence of Off-Target Mutations in the Offspring of Genetically Edited Goats
	Li, Chao 1; Zhou, Shiwei 1; Li, Yan 1; Li, Guanwei 1; Ding, Yige 1; Li, Lan 1; Liu, Jing 1; Qu, Lei 2; Sonstegard, Tad 3; Huang, Xingxu4 ; Jiang, Yu 1; Chen, Yulin 1; Petersen, Bjoern 5; Wang, Xiaolong 1
	2018-10-04
发表期刊	FRONTIERS IN GENETICS (IF:2.8[JCR-2023],3.3[5-Year])
ISSN	1664-8021
卷号	9
发表状态	已发表
DOI	10.3389/fgene.2018.00449
摘要	Unintended off-target mutations induced by CRISPR/Cas9 nucleases may result in unwanted consequences, which will impede the efficient applicability of this technology for genetic improvement. We have recently edited the goat genome through CRISPR/Cas9 by targeting MSTN and FGF5, which increased muscle fiber diameter and hair fiber length, respectively. Using family trio-based sequencing that allow better discrimination of variant origins, we herein generated offspring from edited goats, and sequenced the members of four family trios (gene-edited goats and their offspring) to an average of similar to 36.8x coverage. This data was to systematically examined for mutation profiles using a stringent pipeline that comprehensively analyzed the sequence data for de novo single nucleotide variants, indels, and structural variants from the genome. Our results revealed that the incidence of de novo mutations in the offspring was equivalent to normal populations. We further conducted RNA sequencing using muscle and skin tissues from the offspring and control animals, the differentially expressed genes (DEGs) were related to muscle fiber development in muscles, skin development, and immune responses in skin tissues. Furthermore, in contrast to recently reports of Cas9 triggered p53 expression alterations in cultured cells, we provide primary evidence to show that Cas9-mediated genetic modification does not induce apparent p53 expression changes in animal tissues. This work provides adequate molecular evidence to support the reliability of conducting Cas9-mediated genome editing in large animal models for biomedicine and agriculture.
关键词	genome editing CRISPR/Cas9 whole genome sequencing off-target de novo mutation
收录类别	SCI ; SCIE
语种	英语
资助项目	[2018KJXX-009]
WOS研究方向	Genetics & Heredity
WOS类目	Genetics & Heredity
WOS记录号	WOS:000446361200001
出版者	FRONTIERS MEDIA SA
WOS关键词	RNA-GUIDED ENDONUCLEASES ; CRISPR-CAS NUCLEASES ; HAIR-GROWTH ; HUMAN-CELLS ; GENOME ; FRAMEWORK ; SPECIFICITY ; MUTAGENESIS ; GENERATION ; ALGORITHM
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/28079
专题	生命科学与技术学院_PI研究组_黄行许组
通讯作者	Petersen, Bjoern; Wang, Xiaolong
作者单位	1.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling, Shaanxi, Peoples R China 2.Yulin Univ, Life Sci Res Ctr, Yulin, Peoples R China 3.Recombinetics, St Paul, MN USA 4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 5.Friedrich Loeffler Inst, Inst Nutztiergenet, Neustadt, Germany
推荐引用方式 GB/T 7714	Li, Chao,Zhou, Shiwei,Li, Yan,et al. Trio-Based Deep Sequencing Reveals a Low Incidence of Off-Target Mutations in the Offspring of Genetically Edited Goats[J]. FRONTIERS IN GENETICS,2018,9.
APA	Li, Chao.,Zhou, Shiwei.,Li, Yan.,Li, Guanwei.,Ding, Yige.,...&Wang, Xiaolong.(2018).Trio-Based Deep Sequencing Reveals a Low Incidence of Off-Target Mutations in the Offspring of Genetically Edited Goats.FRONTIERS IN GENETICS,9.
MLA	Li, Chao,et al."Trio-Based Deep Sequencing Reveals a Low Incidence of Off-Target Mutations in the Offspring of Genetically Edited Goats".FRONTIERS IN GENETICS 9(2018).