Knocking-in of the human thrombopoietin gene on beta-casein locus in bovine fibroblasts

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Title
Knocking-in of the human thrombopoietin gene on beta-casein locus in bovine fibroblasts
Author(s)
M Chang; Jeong Woong Lee; D B Koo; S T Shin; Y M Han
Bibliographic Citation
Asian-Australasian Journal of Animal Sciences, vol. 23, no. 6, pp. 806-813
Publication Year
2010
Abstract
Animal bioreactors have been regarded as alternative tools for the production of limited human therapeutic proteins. The mammary glands of cattle are optimal tissues to produce therapeutic proteins that cannot be produced in large amounts in traditional systems based on microorganisms and eukaryotic cells. In this study, two knock-in vectors, pBCTPOKI-6 and pBCTPOKI-10, which target the hTPO gene on the bovine beta-casein locus, were designed to develop cloned transgenic cattle. The pBCTPOKI-6 and pBCTPOKI-10 vectors expressed hTPO protein in culture medium at a concentration of 774 pg/ml and 1,867 pg/ml, respectively. Successfully, two targeted ceil clones were obtained from the bovine fibroblasts transfected with the pBCTPOKI-6 vector. Cloned embryos reconstructed with the targeted nuclei showed a lower in vitro developmental competence than those with the wild-type nuclei. After transfer of the cloned embryos into recipients, 7 pregnancies were detected at 40 to 60 days of gestation, but failed to develop to term. The results are the first trial for targeting of a human gene on the bovine milk protein gene locus, providing the potential for a large-scale production of therapeutic proteins in the animal bioreactor system.
Keyword
Animal bioreactorBovine beta-casein geneBovine fibroblastsHuman thrombopoietin (htpo) geneKnock-inSomatic cell nuclear transfer (SCNT)
ISSN
1011-2367
Publisher
Asian-Australasian Assoc Animal Production Soc
DOI
http://dx.doi.org/10.5713/ajas.2010.90501
Type
Article
Appears in Collections:
Division of Biomedical Research > Biotherapeutics Translational Research Center > 1. Journal Articles
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