title: Generation of germline ablated male pigs by CRISPR/Cas9 editing of the NANOS2 gene.
abstract: Genome editing tools have revolutionized the generation of genetically modified animals including livestock. In particular, the domestic pig is a proven model of human physiology and an agriculturally important species. In this study, we utilized the CRISPR/Cas9 system to edit the NANOS2 gene in pig embryos to generate offspring with mono-allelic and bi-allelic mutations. We found that NANOS2 knockout pigs phenocopy knockout mice with male specific germline ablation but other aspects of testicular development are normal. Moreover, male pigs with one intact NANOS2 allele and female knockout pigs are fertile. From an agriculture perspective, NANOS2 knockout male pigs are expected to serve as an ideal surrogate for transplantation of donor spermatogonial stem cells to expand the availability of gametes from genetically desirable sires.
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.|Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.|Renovate Biosciences Inc, Riesterstown, MD, USA.
authors.forename: Ki-Eun
authors.initials: KE
authors.lastname: Park
authors.affiliation: Center for Reproductive Biology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
authors.forename: Amy V
authors.initials: AV
authors.lastname: Kaucher
authors.affiliation: Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.
authors.forename: Anne
authors.initials: A
authors.lastname: Powell
authors.affiliation: Center for Reproductive Biology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
authors.forename: Muhammad Salman
authors.initials: MS
authors.lastname: Waqas
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.|Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.
authors.forename: Shelley E S
authors.initials: SE
authors.lastname: Sandmaier
authors.affiliation: Center for Reproductive Biology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
authors.forename: Melissa J
authors.initials: MJ
authors.lastname: Oatley
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.|Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.
authors.forename: Chi-Hun
authors.initials: CH
authors.lastname: Park
authors.affiliation: Center for Reproductive Biology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Tibary
authors.affiliation: Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Donovan
authors.affiliation: Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.
authors.forename: Le Ann
authors.initials: LA
authors.lastname: Blomberg
authors.affiliation: Roslin Institute, University of Edinburgh, Edinburgh, Scotland.
authors.forename: Simon G
authors.initials: SG
authors.lastname: Lillico
authors.affiliation: Roslin Institute, University of Edinburgh, Edinburgh, Scotland.
authors.forename: C Bruce A
authors.initials: CB
authors.lastname: Whitelaw
authors.affiliation: Genus PIC, De Forest, WI, USA.
authors.forename: Alan
authors.initials: A
authors.lastname: Mileham
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.|Animal Bioscience and Biotechnology Laboratory, USDA, ARS, Beltsville, MD 20705, USA.|Renovate Biosciences Inc, Riesterstown, MD, USA.
authors.forename: Bhanu P
authors.initials: BP
authors.lastname: Telugu
authors.affiliation: Center for Reproductive Biology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA.
authors.forename: Jon M
authors.initials: JM
authors.lastname: Oatley
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D064113:CRISPR-Cas Systems; D005298:Fertility; D055786:Gene Knockout Techniques; D007248:Infertility, Male; D008297:Male; D016601:RNA-Binding Proteins / Q000235:genetics*; D034421:Sus scrofa / Q000235:genetics*
chemical_list: D016601:RNA-Binding Proteins