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Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk.
Mol Med Rep. 2013 Feb; 7(2):406-12.MM

Abstract

Human interferon α (IFN-α) and erythropoietin (EPO) have been used for a variety of purposes in clinical medicine. Human IFN-α has been used to treat several types of viral infection and cancer, as well as renal anemia, via stimulation of erythrocyte formation in the bone marrow. Transgenic cattle are excellent candidates for pharmaceutical production for humans due to their ability to produce recombinant proteins in milk. The purpose of the present study was to generate bovine transgenic fibroblasts capable of producing recombinant human IFN-α and EPO proteins in transgenic cattle milk. First, we analyzed the promoter activities of various bovine milk protein genes in HC11 mouse mammary epithelial cells. The bovine milk protein gene promoters were cloned into the Luc gene in a promoter-less pGL3-Basic vector. Presence of the αS1-casein promoter (-175 to +796 nt) resulted in an up to 16-fold increase in luciferase activity compared with that of the promoter-less construct. In addition, the human IFN-α and EPO genes were identified as significantly overexpressed in HC11 cells compared with the promoter-less construct. Together, the present results demonstrate that the construct with the αS1-casein promoter may induce secretion of recombinant human IFN-α and EPO into bovine milk. Furthermore, we generated transgenic fibroblasts expressing human IFN-α and EPO cDNA controlled by the αS1-casein promoter and two screening markers, enhanced green fluorescent protein and neomycin resistance. These transgenic fibroblasts may be a source of somatic cells for generating transgenic cattle that produce recombinant human IFN-α and EPO proteins during lactation.

Authors+Show Affiliations

Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23151730

Citation

Jung, Eui-Man, et al. "Establishment of Transgenic Fibroblasts for Producing Recombinant Human Interferon-α and Erythropoietin in Bovine Milk." Molecular Medicine Reports, vol. 7, no. 2, 2013, pp. 406-12.
Jung EM, An BS, Kim YK, et al. Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Mol Med Rep. 2013;7(2):406-12.
Jung, E. M., An, B. S., Kim, Y. K., Hwang, I., Lee, J. Y., Shin, T. Y., Hyun, S. H., Hwang, W. S., & Jeung, E. B. (2013). Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. Molecular Medicine Reports, 7(2), 406-12. https://doi.org/10.3892/mmr.2012.1182
Jung EM, et al. Establishment of Transgenic Fibroblasts for Producing Recombinant Human Interferon-α and Erythropoietin in Bovine Milk. Mol Med Rep. 2013;7(2):406-12. PubMed PMID: 23151730.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Establishment of transgenic fibroblasts for producing recombinant human interferon-α and erythropoietin in bovine milk. AU - Jung,Eui-Man, AU - An,Beum-Soo, AU - Kim,Yu-Kyung, AU - Hwang,Inho, AU - Lee,Jong-Yun, AU - Shin,Tae-Young, AU - Hyun,Sang-Hwan, AU - Hwang,Woo-Suk, AU - Jeung,Eui-Bae, Y1 - 2012/11/13/ PY - 2012/07/18/received PY - 2012/11/07/accepted PY - 2012/11/16/entrez PY - 2012/11/16/pubmed PY - 2013/11/5/medline SP - 406 EP - 12 JF - Molecular medicine reports JO - Mol Med Rep VL - 7 IS - 2 N2 - Human interferon α (IFN-α) and erythropoietin (EPO) have been used for a variety of purposes in clinical medicine. Human IFN-α has been used to treat several types of viral infection and cancer, as well as renal anemia, via stimulation of erythrocyte formation in the bone marrow. Transgenic cattle are excellent candidates for pharmaceutical production for humans due to their ability to produce recombinant proteins in milk. The purpose of the present study was to generate bovine transgenic fibroblasts capable of producing recombinant human IFN-α and EPO proteins in transgenic cattle milk. First, we analyzed the promoter activities of various bovine milk protein genes in HC11 mouse mammary epithelial cells. The bovine milk protein gene promoters were cloned into the Luc gene in a promoter-less pGL3-Basic vector. Presence of the αS1-casein promoter (-175 to +796 nt) resulted in an up to 16-fold increase in luciferase activity compared with that of the promoter-less construct. In addition, the human IFN-α and EPO genes were identified as significantly overexpressed in HC11 cells compared with the promoter-less construct. Together, the present results demonstrate that the construct with the αS1-casein promoter may induce secretion of recombinant human IFN-α and EPO into bovine milk. Furthermore, we generated transgenic fibroblasts expressing human IFN-α and EPO cDNA controlled by the αS1-casein promoter and two screening markers, enhanced green fluorescent protein and neomycin resistance. These transgenic fibroblasts may be a source of somatic cells for generating transgenic cattle that produce recombinant human IFN-α and EPO proteins during lactation. SN - 1791-3004 UR - https://www.unboundmedicine.com/medline/citation/23151730/Establishment_of_transgenic_fibroblasts_for_producing_recombinant_human_interferon_α_and_erythropoietin_in_bovine_milk_ L2 - http://www.spandidos-publications.com/mmr/7/2/406 DB - PRIME DP - Unbound Medicine ER -