Dominant-negative mutant hepatocyte nuclear factor 1α induces diabetes in transgenic-cloned pigs

Abstract

Pigs have been recognized as an excellent biomedical model for investigating a variety of human health issues. We developed genetically modified pigs that exhibit the apparent symptoms of diabetes. Transgenic cloned pigs carrying a mutant human hepatocyte nuclear factor 1alpha gene, which is known to cause the type 3 form of maturity-onset diabetes of the young, were produced using a combined technology of intracytoplasmic sperm injection-mediated gene transfer and somatic cell nuclear transfer. Although most of the 22 cloned offspring obtained died before weaning, four pigs that lived for 20-196 days were diagnosed as diabetes mellitus with nonfasting blood glucose levels greater than 200 mg/dl. Oral glucose tolerance test on a cloned pig also revealed a significant increase of blood glucose level after glucose loading. Histochemical analysis of pancreas tissue from the cloned pigs showed small and irregularly formed Langerhans Islets, in which poor insulin secretion was detected.

Livestock Production via Micromanipulation

“…This technique is advantageous in the stable incorporation and expression of large (>100 kb) DNA constructs such as yeast artifi cial chromosomes (YACs) that are not amenable to transfection or viral delivery [ 18 ] . The successful production of nontargeted transgenic pigs following both ICSI and NT techniques has been reported, albeit without using artifi cial chromosomes [ 19 ] .…”

39 Factors Influencing the Efficiency of Generating Genetically Engineered Pigs by Nuclear Transfer: Multi-Factorial Analysis of a Large Data Set

“…Genetically tailored pig models have already been developed to investigate cystic fibrosis [12], diabetes mellitus [13][14][15][16], and neurodegenerative diseases [17] (reviewed in [18]). Multiple lines of genetically modified pigs have also been generated for xenotransplantation (reviewed in [19]), most notably α1,3-galactosyl transferase knockout pigs lacking α1,3-Gal, the major xeno-antigen [5].…”

Cloned Porcine Embryos can Maintain Developmental Ability after Cryopreservation at the Morula Stage

“…NAKANO et al 314 preservation were transferred at the early cleavage stage (days 1-2), avoiding a prolonged culture that may have an adverse effect on the viability of embryos. This timing for transfer of the noncryopreserved SCNT embryos is routinely used as the most feasible condition for cloned pig production in our laboratory [14,15,21,23]. Furthermore, noncryopreserved embryos were transferred to sexually mature pregnant pigs that had been artificially aborted, while gilts were used as the recipients for transferring the cryopreserved embryos.…”

“…57: 312-316, 2011) n recent years, pigs have often been used as large laboratory animals in biomedical research [1][2][3][4][5][6][7], and genetically engineered pigs are highly desired in rapidly advancing research areas, such as regenerative medicine, organ transplantation, analysis of intractable genetic disorders and stem cell therapy [for review, see 7]. For the production of genetically engineered pigs, including gene knockout pigs, somatic cell cloning is a key technology [8][9][10][11][12][13][14][15]. Therefore, if a reliable cryopreservation technique for cloned embryos can be established, the production and application of various specially designed pigs will advance further.…”