Recombinant human erythropoietin produced in milk of transgenic pigs

Park, Jin-Ki; Lee, Yun-Keun; Lee, Poongyeon; Chung, Hak-Jae; Kim, SungWoo; Lee, Hyun-Gi; Seo, Myung-Kyu; Han, Joo-Hee; Park, Chun-Gyu; Kim, Hun-Taek; Kim, Yongkook; Min, Kwan-Sik; Kim, Jin‐Hoi; Lee, Hoon-Taek; Chang, Won-Kyong

doi:10.1016/j.jbiotec.2005.11.021

Cited by 38 publications

(24 citation statements)

References 45 publications

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“…Using various mammary gland-specific or blood-specific promoters to drive the expression of specific protein-coding genes, transgenic domestic animals can continuously produce the recombinant proteins in large quantities in their milk or blood. Recombinant proteins, including human von Willebrand factor (Lee et al, 2009), human erythropoietin (Park et al, 2006), human insulin-like growth factor-I (Monaco et al, 2005), human factor VIII (Paleyanda et al, 1997) and bovine alpha-lactalbumin (Bleck et al, 1998) have been produced in the milk of transgenic pigs. Transgenic goats, capable of synthesizing human butyrylcholinesterase (Huang et al, 2007) and human longer acting tissue plasminogen activator (Ebert et al, 1991) in their milk have also been created.…”

Section: Production Of Recombinant Proteinsmentioning

confidence: 99%

Genetic Modification of Domestic Animals for Agriculture and Biomedical Applications

Yang¹,

Ross²

2012

Biomedical Science, Engineering and Technology

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Section: Production Of Recombinant Proteinsmentioning

confidence: 99%

Genetic Modification of Domestic Animals for Agriculture and Biomedical Applications

Yang¹,

Ross²

2012

Biomedical Science, Engineering and Technology

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“…More recently, transgenic pigs were generated for the production of recombinant EPO in their milk. However, transgenic EPO expression resulted in high embryonic mortality, in addition to sterility [12].…”

Section: Introductionmentioning

confidence: 99%

Molecular Cloning and Transgenic Expression of a Synthetic Human Erythropoietin Gene in Tobacco

Sperb

Werlang

Margis‐Pinheiro

et al. 2011

Appl Biochem Biotechnol

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Erythropoietin (EPO) is a hormone belonging to a group of hematopoietic growth factors that control the proliferation and differentiation of bone marrow cells. It induces the production of erythrocytes, thereby increasing the amount of circulating hemoglobin and oxygen. Previous attempts to transgenically express human EPO in plants failed to succeed because the plants exhibited abnormal morphology and infertility. In the present work, we describe the generation of fertile transgenic tobacco plants able to express a synthetic version of human EPO. A 582-bp fragment of the human EPO gene was synthesized using a PCR-based method and ligated into pCR-Blunt. After sequencing, the human EPO fragment was transferred to pWUbi.tm1 and the expression cassette was then transferred to the binary vector pWBVec4a. After Agrobacterium-mediated transformation of Nicotiana tabacum SR1 plants, integration of the transgene into T(0) and T(1) plant genomes was confirmed by PCR. The human EPO gene was found to be expressed in tobacco leaves at the mRNA and protein levels. Self-crossing allowed us to obtain T(1) plants exhibiting Mendelian segregation of the transgene. None of the plants presented any kind of malformation or deformity.

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“…Gordon et al were the first to attempt to produce recombinant proteins in the milk of transgenic (TG) mice [6]; since then, modification of phenotypic properties became possible [7]. Of the transgenic technologies employing domestic animals that have been developed over the last decade, one of the most popular applications has been the generation of animal bioreactors for production of therapeutic proteins [8][9][10][11][12]. The pig is an attractive animal for this kind of application due to its fecundity and physiology that closely resembles that of humans [13].…”

mentioning

confidence: 99%

“…Mass production of therapeutic proteins in the milk of TG pigs is a cost-effective alternative to production by cell culture or fermentation [14]. There have been several reports on use of the pig as an animal bioreactor for recombinant proteins such as human FVIII [15], human hemoglobin [16], human protein C [17] and human erythropoietin [9]. Despite the usefulness of this technique for analysis of protein functions and development of model animals for studying diseases, there have been only a few reports on largescale production of TG pigs expressing human proteins.…”

mentioning

confidence: 99%

Production of Recombinant Human Von Willebrand Factor in the Milk of Transgenic Pigs

Lee

Kim

et al. 2009

Journal of Reproduction and Development

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Abstract. Von Willebrand factor (vWF), a large multimeric glycoprotein present in blood plasma, is a blood protein of the coagulation system. It is defective in von Willebrand disease and is involved in a large number of other diseases, including thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and heyde's syndrome. We have developed a line of transgenic swine harboring recombinant human von Willebrand factor (rhvWF) cDNA through microinjection of fertilized one-cell pig zygotes. Expression of rhvWF in the mammary gland and secretion of rhvWF into the milk of the transgenic swine were confirmed by immunohistochemical and western blot analyses, respectively, and rhvWF proteins were detected in milk from all lactating founder females at concentrations that were 28-to 56-folds greater than that in circulating human plasma. The amino acid sequence of rhvWF protein in the transgenic pig milk matched that of vWF produced from human blood plasma. This study provides evidence that production of rhvWF from transgenic pig milk is a potentially valuable technology and can be used as a cost-effective alternative in clinical applications. on Willebrand factor (vWF or factor VIII-related antigen)-the deficiency of which causes von Willebrand disease (vWD), the most common inherited bleeding disorder-is constitutively produced in the endothelium, megakaryocytes and subendothelial connective tissue as a large multimeric glycoprotein circulating in blood plasma [1,2]. It is also involved in a large number of other diseases, including thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and heyde's syndrome [3,4], mediates platelet adhesion to sites of vascular injury by binding to specific platelet membrane glycoproteins in a closely regulated manner and functions as a carrier protein for blood clotting factor VIII (FVIII) [5]. In the circulation, vWF mainly functions by binding to other proteins, particularly FVIII, during the process of coagulation, and the absence of this interaction causes rapid degradation of vWF. In regard to the apparent role of vWF in high shear stress, vWD deficiency leads to a tendency to bleed, most apparently in tissues with high blood flow in narrow vessels [2]. The basic vWF monomer is a 2,050 amino acid protein and is assembled via identical 250-kDa subunits into disulfide-linked multimers that may vary between 2 and >20; thus, the molecular mass of the heterogeneously sized vWF protein circulating in plasma ranges from 5 × 10 5 Da to more than 5 × 10 6 Da [1, 2]. The development of a method for production of a highly purified protein on an economically large scale is one of the targets of modern animal biotechnology research. Gordon et al. were the first to attempt to produce recombinant proteins in the milk of transgenic (TG) mice [6]; since then, modification of phenotypic properties became possible [7]. Of the transgenic technologies employing domestic animals that have been developed over the last decade, one of the most popular applications has been the generation of an...

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Recombinant human erythropoietin produced in milk of transgenic pigs

Cited by 38 publications

References 45 publications

Genetic Modification of Domestic Animals for Agriculture and Biomedical Applications

Genetic Modification of Domestic Animals for Agriculture and Biomedical Applications

Molecular Cloning and Transgenic Expression of a Synthetic Human Erythropoietin Gene in Tobacco

Production of Recombinant Human Von Willebrand Factor in the Milk of Transgenic Pigs

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