title: Disruption of FGF5 in Cashmere Goats Using CRISPR/Cas9 Results in More Secondary Hair Follicles and Longer Fibers.
abstract: Precision genetic engineering accelerates the genetic improvement of livestock for agriculture and biomedicine. We have recently reported our success in producing gene-modified goats using the CRISPR/Cas9 system through microinjection of Cas9 mRNA and sgRNAs targeting the MSTN and FGF5 genes in goat embryos. By investigating the influence of gene modification on the phenotypes of Cas9-mediated goats, we herein demonstrate that the utility of this approach involving the disruption of FGF5 results in increased number of second hair follicles and enhanced fiber length in Cas9-mediated goats, suggesting more cashmere will be produced. The effects of genome modifications were characterized using H&E and immunohistochemistry staining, quantitative PCR, and western blotting techniques. These results indicated that the gene modifications induced by the disruption of FGF5 had occurred at the morphological and genetic levels. We further show that the knockout alleles were likely capable of germline transmission, which is essential for goat population expansion. These results provide sufficient evidences of the merit of using the CRISPR/Cas9 approach for the generation of gene-modified goats displaying the corresponding mutant phenotypes.
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: Bei
authors.initials: B
authors.lastname: Cai
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China.|School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
authors.forename: Jiankui
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin 719000, China.|Life Science Research Center, Yulin University, Yulin 719000, China.
authors.forename: Haijing
authors.initials: H
authors.lastname: Zhu
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: Yiyuan
authors.initials: Y
authors.lastname: Niu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Baohua
authors.initials: B
authors.lastname: Ma
authors.affiliation: Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin 719000, China.|Life Science Research Center, Yulin University, Yulin 719000, China.
authors.forename: Honghao
authors.initials: H
authors.lastname: Yu
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Anmin
authors.initials: A
authors.lastname: Lei
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.|Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin 719000, China.|Life Science Research Center, Yulin University, Yulin 719000, China.
authors.forename: Hailong
authors.initials: H
authors.lastname: Yan
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Qiaoyan
authors.initials: Q
authors.lastname: Shen
authors.affiliation: Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin 719000, China.|Life Science Research Center, Yulin University, Yulin 719000, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Xiaoe
authors.initials: X
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Jinlian
authors.initials: J
authors.lastname: Hua
authors.affiliation: MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, National Resource Center for Mutant Mice, Nanjing 210061, China.|School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
authors.forename: Xingxu
authors.initials: X
authors.lastname: Huang
authors.affiliation: Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin 719000, China.|Life Science Research Center, Yulin University, Yulin 719000, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Qu
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D051521:Fibroblast Growth Factor 5 / Q000737:chemistry / Q000172:deficiency / Q000235:genetics*; D018095:Germ-Line Mutation; D006041:Goats; D018859:Hair Follicle / Q000737:chemistry* / Q000473:pathology / Q000502:physiology; D008297:Male; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D012867:Skin / Q000473:pathology; D013737:Testis / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D051521:Fibroblast Growth Factor 5