Base pair editing in goat: nonsense codon introgression into FGF5 results in longer hair

doi:10.1111/febs.14983

	Base pair editing in goat: nonsense codon introgression into FGF5 results in longer hair
	Li, Guanwei 1; Zhou, Shiwei 1; Li, Chao 1; Cai, Bei 1; Yu, Honghao 2; Ma, Baohua 3; Huang, Yu 1; Ding, Yige 1; Liu, Yao 1; Ding, Qiang 1; He, Chong 4; Zhou, Jiankui5 ; Wang, Ying 1; Zhou, Guangxian 1; Li, Yan 1; Yan, Yuan 3; Hua, Jinlian 3; Petersen, Bjoern 6; Jiang, Yu 1; Sonstegard, Tad 7; Huang, Xingxu5 ; Chen, Yulin 1; Wang, Xiaolong 1
	2019-07-23
发表期刊	FEBS JOURNAL (IF:5.5[JCR-2023],5.5[5-Year])
ISSN	1742-464X
发表状态	已发表
DOI	10.1111/febs.14983
摘要	The ability to alter single bases without homology directed repair (HDR) of double-strand breaks provides a potential solution for editing livestock genomes for economic traits, which are often multigenic. Progress toward multiplex editing in large animals has been hampered by the costly inefficiencies of HDR via microinjection of in vitro manipulated embryos. Here, we designed sgRNAs to induce nonsense codons (C-to-T transitions) at four target sites in caprine FGF5, which is a crucial regulator of hair length in mammals. Initial transfections of the third generation Base Editor (BE3) plasmid and four different sgRNAs into caprine fibroblasts were ineffective in altering FGF5. In contrast, all five progenies produced from microinjected single-cell embryos had alleles with a targeted nonsense mutation. The effectiveness of BE3 to make single base changes varied considerably based on sgRNA design. In addition, the rate of mosaicism differed between animals, target sites, and tissue type. The phenotypic effects on hair fiber were characterized by hematoxylin and eosin, immunofluorescence staining, and western blotting. Differences in morphology were detectable, even though mosaicism was probably affecting the levels of FGF5 expression. PCR amplicon and whole-genome resequencing analyses for off-target changes caused by BE3 were low at a genome-wide scale. This study provided the first evidence of base editing in large mammals produced from microinjected single-cell embryos. Our results support further optimization of BEs for introgressing complex human disease alleles into large animal models, to evaluate potential genetic improvement of complex health and production traits in a single generation.
关键词	base editing genome editing large animal nonsense mutation off-target mutation point mutation
收录类别	SCI ; SCIE
语种	英语
资助项目	China Agriculture Research System[CARS-39]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000477467500001
出版者	WILEY
WOS关键词	ONE-STEP GENERATION ; EFFICIENT GENERATION ; READ ALIGNMENT ; KNOCKOUT MICE ; TARGET BASE ; GENE ; MUTATIONS ; RICE ; REGULATOR ; ALLELES
原始文献类型	Article ; Early Access
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/61058
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; Chen, Yulin; Wang, Xiaolong
作者单位	1.Northwest A&F Univ, Coll Anim Sci & Technol, Key Lab Anim Genet Breeding & Reprod Shaanxi Prov, Yangling 712100, Shaanxi, Peoples R China 2.Guilin Med Univ, Coll Biotechnol, Guilin, Peoples R China 3.Northwest A&F Univ, Coll Vet Med, Yangling, Shaanxi, Peoples R China 4.Northwest A&F Univ, Coll Informat & Engn, Yangling, Shaanxi, Peoples R China 5.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 6.Friedrich Loeffler Inst, Inst Farm Anim Genet, Neustadt, Germany 7.Recombinetics, St Paul, MN USA
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Li, Guanwei,Zhou, Shiwei,Li, Chao,et al. Base pair editing in goat: nonsense codon introgression into FGF5 results in longer hair[J]. FEBS JOURNAL,2019.
APA	Li, Guanwei.,Zhou, Shiwei.,Li, Chao.,Cai, Bei.,Yu, Honghao.,...&Wang, Xiaolong.(2019).Base pair editing in goat: nonsense codon introgression into FGF5 results in longer hair.FEBS JOURNAL.
MLA	Li, Guanwei,et al."Base pair editing in goat: nonsense codon introgression into FGF5 results in longer hair".FEBS JOURNAL (2019).