title: Establishment of a canine model of human type 2 diabetes mellitus by overexpressing phosphoenolypyruvate carboxykinase.
abstract: Dogs are useful models for studying human metabolic diseases such as type 2 diabetes mellitus due to similarities in physiology, anatomy and life styles with humans. Somatic cell nuclear transfer (SCNT) facilitates the production of transgenic dogs. In this study, we generated transgenic dogs expressing the phosphoenolpyruvate carboxykinase (PEPCK) gene, which is closely involved in the pathogenesis of type 2 diabetes mellitus. In addition, we assessed the cloning efficiency associated with adult or fetal (cloned or natural mating) fibroblasts as a nuclear source. Cloning efficiency was determined by the fusion, pregnancy and cloning rates. The fusion rates were significantly high for fibroblasts from cloned fetuses, but the pregnancy and cloning rates were relatively high for cells from normal fetuses. Based on these data, fetal fibroblasts were selected as the nuclear donor for SCNT and genetically engineered to overexpress the PEPCK gene and dual selection marker genes controlled by the PEPCK promoter. The transgenic cells were introduced into oocytes and transferred into five recipient dogs, resulting in two pregnancies. Finally, three puppies were born and confirmed by microsatellite analysis to be genetically identical to the donor. One puppy successfully overexpressed PEPCK mRNA and protein in the liver. This canine disease model may be useful for studying the pathogenesis and/or therapeutic targets of type 2 diabetes mellitus.
authors.affiliation: SooAm Biotech Research Foundation, Seoul 152-904, Republic of                     Korea.
authors.forename: Yeon Woo
authors.initials: YW
authors.lastname: Jeong
authors.forename: Geun-Shik
authors.initials: GS
authors.lastname: Lee
authors.forename: Joung Joo
authors.initials: JJ
authors.lastname: Kim
authors.forename: Sun Woo
authors.initials: SW
authors.lastname: Park
authors.forename: Kyeong Hee
authors.initials: KH
authors.lastname: Ko
authors.forename: Mina
authors.initials: M
authors.lastname: Kang
authors.forename: Yu Kyung
authors.initials: YK
authors.lastname: Kim
authors.forename: Eui-Man
authors.initials: EM
authors.lastname: Jung
authors.forename: Sang Hwan
authors.initials: SH
authors.lastname: Hyun
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Shin
authors.forename: Eui-Bae
authors.initials: EB
authors.lastname: Jeung
authors.forename: Woo Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D003924:Diabetes Mellitus, Type 2 / Q000201:enzymology / Q000235:genetics*; D004195:Disease Models, Animal; D004285:Dogs / Q000235:genetics; D004624:Embryo Transfer / Q000379:methods; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D023061:Gene Order; D018390:Gene Targeting; D005822:Genetic Vectors; D006801:Humans; D053652:Nuclear Transfer Techniques; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism
chemical_list: C069767:phosphoenolpyruvate carboxylase kinase; D017346:Protein Serine-Threonine Kinases