title: Efficient gene knockout in goats using CRISPR/Cas9 system.
abstract: The CRISPR/Cas9 system has been adapted as an efficient genome editing tool in laboratory animals such as mice, rats, zebrafish and pigs. Here, we report that CRISPR/Cas9 mediated approach can efficiently induce monoallelic and biallelic gene knockout in goat primary fibroblasts. Four genes were disrupted simultaneously in goat fibroblasts by CRISPR/Cas9-mediated genome editing. The single-gene knockout fibroblasts were successfully used for somatic cell nuclear transfer (SCNT) and resulted in live-born goats harboring biallelic mutations. The CRISPR/Cas9 system represents a highly effective and facile platform for targeted editing of large animal genomes, which can be broadly applied to both biomedical and agricultural applications.
authors.affiliation: College of Life Sciences, Shihezi University, Shihezi, Xinjiang, China; Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Qiao
authors.affiliation: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
authors.forename: Shengwei
authors.initials: S
authors.lastname: Hu
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang, China.
authors.forename: Xinxia
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
authors.forename: Misha
authors.initials: M
authors.lastname: Regouski
authors.affiliation: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, United States of America.
authors.forename: Irina A
authors.initials: IA
authors.lastname: Polejaeva
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang, China.
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
mesh_terms: D000483:Alleles; D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D004624:Embryo Transfer; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D055786:Gene Knockout Techniques; D016678:Genome; D006041:Goats / Q000235:genetics*; D007782:Lactoglobulins / Q000172:deficiency / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D055435:Myostatin / Q000172:deficiency / Q000235:genetics; D028861:Nuclear Pore Complex Proteins / Q000172:deficiency / Q000235:genetics; D053652:Nuclear Transfer Techniques; D012150:Polymorphism, Restriction Fragment Length; D061251:Primary Cell Culture; D011328:Prions / Q000037:antagonists & inhibitors / Q000235:genetics
chemical_list: D007782:Lactoglobulins; D055435:Myostatin; D028861:Nuclear Pore Complex Proteins; D011328:Prions