Cloned Transgenic Calves Produced from Nonquiescent Fetal Fibroblasts

Cibelli, José B.; Stice, S.L.; Golueke, Paul J.; Kane, Jeff; Jerry, Joseph; Blackwell, Catherine; León, F. A. Ponce de; Robl, James M.

doi:10.1126/science.280.5367.1256

Cited by 1,297 publications

(770 citation statements)

References 11 publications

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“…Gene targeting has many useful applications in science, medicine and industry and may be one of the most useful applications of somatic cell cloning technology. Currently, gene targeting using ES cells has been successful only in mice, but somatic cell cloning has been successful for many species [21][22][23][24][25] . The results obtained in this study indicate that complex genetic modifications can now be readily made for a wide variety of genes in many species.…”

Section: Discussionmentioning

confidence: 99%

Sequential targeting of the genes encoding immunoglobulin-μ and prion protein in cattle

et al. 2004

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Gene targeting is accomplished using embryonic stem cells in the mouse but has been successful, only using primary somatic cells followed by embryonic cloning, in other species. Gene targeting in somatic cells versus embryonic stem cells is a challenge; consequently, there are few reported successes and none include the targeting of transcriptionally silent genes or double targeting to produce homozygotes. Here, we report a sequential gene targeting system for primary fibroblast cells that we used to knock out both alleles of a silent gene, the bovine gene encoding immunoglobulin-µ (IGHM), and produce both heterozygous and homozygous knockout calves. We also carried out sequential knockout targeting of both alleles of a gene that is active in fibroblasts, encoding the bovine prion protein (PRNP), in the same genetic line to produce doubly homozygous knockout fetuses. The sequential gene targeting system we used alleviates the need for germline transmission for complex genetic modifications and should be broadly applicable to gene functional analysis and to biomedical and agricultural applications.

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Section: Discussionmentioning

confidence: 99%

Sequential targeting of the genes encoding immunoglobulin-μ and prion protein in cattle

et al. 2004

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“…With the successful cloning of animals by somatic cell nuclear transfer (SCNT), it is now possible to produce transgenic pigs from genetically engineered somatic donor cells. In SCNT, reported for a variety of animal species including the mouse (Wakayama et al 2000; Yanagimachi 1999), the sheep (Wilmut et al 1997), the cow (Cibelli et al 1998;Kato et al 1998;Kubota et al 2000), and the pig (Betthauser et al 2000;Onishi et al 2000;Polejaeva et al 2000), the nucleus from a single differentiated somatic cell is transferred into an enucleated oocyte (unfertilized egg cell), and the reconstructed embryo is subsequently transferred to a surrogate mother. This procedure allows modification of the somatic donor cell in culture by transgene insertion or introduction of loss-offunction gene knockout mutations by homologous recombination.…”

Section: Introductionmentioning

confidence: 99%

Pig transgenesis by Sleeping Beauty DNA transposition

Jakobsen

Kragh

et al. 2010

Transgenic Res

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Modelling of human disease in genetically engineered pigs provides unique possibilities in biomedical research and in studies of disease intervention. Establishment of methodologies that allow efficient gene insertion by non-viral gene carriers is an important step towards development of new disease models. In this report, we present transgenic pigs created by Sleeping Beauty DNA transposition in primary porcine fibroblasts in combination with somatic cell nuclear transfer by handmade cloning. Göttingen minipigs expressing green fluorescent protein are produced by transgenesis with DNA transposon vectors carrying the transgene driven by the human ubiquitin C promoter. These animals carry multiple copies (from 8 to 13) of the transgene and show systemic transgene expression. Transgene-expressing pigs carry both transposase-catalyzed insertions and at least one copy of randomly inserted plasmid DNA. Our findings illustrate critical issues related to DNA transposon-directed transgenesis, including coincidental plasmid insertion and relatively low Sleeping Beauty transposition activity in porcine fibroblasts, but also provide a platform for future development of porcine disease models using the Sleeping Beauty gene insertion technology.

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“…The somatic cloning technology for farm animals is improving, especially in the bovine species. Over 2000 cloned cattle have been obtained worldwide since 1998 when the first calves were born from somatic cell nuclear transfer (Cibelli et al, 1998;Vignon et al, 1998). Before the commercial use of food products derived from cloned animals is possible, an evaluation of the products and an assessment of the possible risks associated with consumption are required.…”

Section: Introductionmentioning

confidence: 99%

Comparison of cloned and non-cloned Holstein heifers in muscle contractile and metabolic characteristics

Jurie

Picard

Heyman

et al. 2009

Animal

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Muscle contractile and metabolic characteristics were studied on nine cloned and eight non-cloned (control) heifers. The animals were submitted to repeated biopsies of the semitendinosus (ST) muscle at the ages of 8, 12, 18 and 24 months. The contractile type was determined from the proportion of the different myosin heavy chain (MyHC) isoforms separated by electrophoresis. Glycolytic metabolism was assessed by lactate dehydrogenase (LDH) activity, and oxidative metabolism was assessed by isocitrate dehydrogenase (ICDH), cytochrome-c oxidase (COX) and b-hydroxyacyl-CoA dehydrogenase (HAD) activities. In cloned heifers at 8 months of age, there was a greater proportion of MyHC I (slow oxidative isoform) and MyHC IIa (fast oxido-glycolytic isoform), a lower proportion of MyHC IIx (fast glycolytic isoform), greater COX and HAD activity and a lower LDH/ICDH ratio compared with control heifers. Thus, young cloned heifers had slower muscle types associated with a more oxidative muscular metabolism than control heifers. From 12 months of age onwards, no significant differences were observed between cloned and control heifers. A delay in muscle differentiation and maturation in cloned heifers is hypothesised and discussed.

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Cloned Transgenic Calves Produced from Nonquiescent Fetal Fibroblasts

Cited by 1,297 publications

References 11 publications

Sequential targeting of the genes encoding immunoglobulin-μ and prion protein in cattle

Sequential targeting of the genes encoding immunoglobulin-μ and prion protein in cattle

Pig transgenesis by Sleeping Beauty DNA transposition

Comparison of cloned and non-cloned Holstein heifers in muscle contractile and metabolic characteristics

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