title: Production of cloned NIBS (Nippon Institute for Biological Science) and α-1, 3-galactosyltransferase knockout MGH miniature pigs by somatic cell nuclear transfer using the NIBS breed as surrogates.
abstract: BACKGROUND
  Nuclear transfer (NT) technologies offer a means for producing the genetically modified pigs necessary to develop swine models for mechanistic studies of disease processes as well as to serve as organ donors for xenotransplantation. Most previous studies have used commercial pigs as surrogates.
  
  
  RESULTS
  In this study, we established a cloning technique for miniature pigs by somatic cell nuclear transfer (SCNT) using Nippon Institute for Biological Science (NIBS) miniature pigs as surrogates. Moreover, utilizing this technique, we have successfully produced an α-1, 3-galactosyltransferase knockout (GalT-KO) miniature swine. Fibroblasts procured from a NIBS miniature pig fetus were injected into 1312 enucleated oocytes. The cloned embryos were transferred to 11 surrogates of which five successfully delivered 13 cloned offspring; the production efficiency was 1.0% (13/1312). In a second experiment, lung fibroblasts obtained from neonatal GalT-KO MGH miniature swine were used as donor cells and 1953 cloned embryos were transferred to 12 surrogates. Six cloned offspring were born from five surrogates, a production efficiency of 0.3% (6/1953).
  
  
  CONCLUSIONS
  These results demonstrate successful establishment of a miniature pig cloning technique by SCNT using NIBS miniature pigs as surrogates. To our knowledge, this is the first demonstration of successful production of GalT-KO miniature swine using miniature swine surrogates. This technique could help to ensure a stable supply of the cloned pigs through the use of miniature pig surrogates and could expand production in countries with limited space or in facilities with special regulations such as specific pathogen-free or good laboratory practice.
authors.affiliation: Research & Development Department, Nippon Institute for Biological Science, Hokuto, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Shimatsu
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Yamada
authors.forename: Wataru
authors.initials: W
authors.lastname: Horii
authors.forename: Atsushi
authors.initials: A
authors.lastname: Hirakata
authors.forename: Yuji
authors.initials: Y
authors.lastname: Sakamoto
authors.forename: Shiori
authors.initials: S
authors.lastname: Waki
authors.forename: Junichi
authors.initials: J
authors.lastname: Sano
authors.forename: Toshiki
authors.initials: T
authors.lastname: Saitoh
authors.forename: Hisashi
authors.initials: H
authors.lastname: Sahara
authors.forename: Akira
authors.initials: A
authors.lastname: Shimizu
authors.forename: Hajime
authors.initials: H
authors.lastname: Yazawa
authors.forename: David H
authors.initials: DH
authors.lastname: Sachs
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nunoya
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D005700:Galactosyltransferases / Q000172:deficiency* / Q000235:genetics* / Q000276:immunology; D055786:Gene Knockout Techniques / Q000662:veterinary; D008297:Male; D053652:Nuclear Transfer Techniques; D011247:Pregnancy; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics* / Q000276:immunology; D014183:Transplantation, Heterologous
chemical_list: D005700:Galactosyltransferases; C530267:alpha-1,3-galactosyltransferase 1, porcine
keywords: GalT-KO MGH miniature; Nippon Institute for Biological Science miniature pigs; embryo transfer; miniature pigs; somatic cell nuclear transfer; α-1,3-galactosyltransferase knockout