High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

Velander, William H.; Johnson, John L.; Page, Raymond; Russell, Christopher G.; Subramanian, Anuradha; Wilkins, Tracy D.; Gwazdauskas, F.C.; Pittius, Christoph W.; Drohan, William N.

doi:10.1073/pnas.89.24.12003

Cited by 150 publications

(66 citation statements)

References 17 publications

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“…For example, the antitrypsin protein was expressed in the transgenic sheep mammary latex, at a maximal concentration of 37.5 g/L, which is over 50% of the total latex protein concentration [39] . The concentrations of the whey acidic protein (WAP) and the human protein C (HPC) in transgenic pig latex were 1 g/L [40,41] . Although the use of bioreactors for protein production has a high societal appeal, there are only a few cases that have gone to Phase II or III clinical trials.…”

Section: Secretion Of Transgenic Spermary-specific Fish Bioreactormentioning

confidence: 99%

A perspective on fish gonad manipulation for biotechnical applications

Wang

Zhang

2006

Chin. Sci. Bull.

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The gonad is an essential organ for generating sperm and ova in vertebrates. This review describes several pilot studies on gonad gene manipulation and development in fish. With antisense RNA techniques, we suppressed the gonad development, and thus the fertility, of an antisense gonadotropin-releasing hormone (sGnRH) transgenic common carp. Then, using a tissue-specific exogenous gene excision strategy with sexual compensation, we knocked out the gonad-specific transgene. Under the control of the rainbow trout protamine promoter, the transgenic fish expressed the reporter gene eGFP specifically in the spermary. These results indicate that the fish gonad is a new model organ that can improve contemporary biotechnology experiments. Herein we discuss the potential of fish gonad manipulation for resolving important biosafety problems regarding transgenic fish generation and producing the new transgenic animal bioreactor.

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Section: Secretion Of Transgenic Spermary-specific Fish Bioreactormentioning

confidence: 99%

A perspective on fish gonad manipulation for biotechnical applications

Wang

Zhang

2006

Chin. Sci. Bull.

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“…For the purposes of pharmaceutical production at large-scale, a single cointegration site and associated transgene copy number in that locus is desirable for the facile establishment of a phenotypical and genotypical stable lineage . Mosaicism and multiple integration sites frequently occur in founder animals and this complicates analysis of founder animals (Velander et al, 1992b). Thus, phenotype and genotype can not be reliably defined in transgenic animals until successive generations of offspring obtained from outbreeding with nontransgenic animals are analyzed Velander et al, 1992b;Carver et al, 1993).…”

Section: Transgenic Livestock As An Alternative Source Of Recombinantmentioning

confidence: 99%

“…Mosaicism and multiple integration sites frequently occur in founder animals and this complicates analysis of founder animals (Velander et al, 1992b). Thus, phenotype and genotype can not be reliably defined in transgenic animals until successive generations of offspring obtained from outbreeding with nontransgenic animals are analyzed Velander et al, 1992b;Carver et al, 1993). A production lineage must be shown to reproducibly secrete the recombinant protein at a level and functionality which can be associated with a given trans-genotypic signature.…”

Section: Transgenic Livestock As An Alternative Source Of Recombinantmentioning

confidence: 99%

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Production and secretion of recombinant human fibrinogen by the transgenic murine mammary gland

Butler¹,

O'Sickey²,

Lord³

et al. 1999

Theriogenology

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The mammary gland of lactating transgenic animals has several advantages for production of heterologous proteins including a high cell density that results in high concentrations of secreted protein and the ability to perform several types of post-translational modifications. Transgenes were constructed from the 4.1 kbp murine Whey Acidic Protein promoter (mWAP) and the three cDNAs coding for the A", B$ and ( fibrinogen chains to evaluate the requirements of the transgenic murine mammary gland for high level secretion of fully assembled human fibrinogen. After introducing the constructs into the murine zygotes by microinjection, secretion of fully assembled fibrinogen into milk was measured at concentrations between 10 :g/ml to 200 :g/ml.In one line of mice the total secretion of fibrinogen and unassembled subunits approached 700 :g/ml in milk. The level of assembled fibrinogen was proportional to the lowest amount of subunit produced where both the B$ and ( chains were rate limiting. Also, the subunit complexes ( 2 , A"( 2 and the individual subunits A", B$ and ( were found as secretion products. This is the first time that secretion of individual B$-subunits by any cell type has been reported and suggests the organization of the secretion pathway in mammary epithelia is different from that in liver. Glycosylated forms of individual B$-chain contained a complex saccharide with low mannose. Glycosylation of the (-chain was also observed. These results suggest the 4.1 mWAP promoter can drive expression of fibrinogen cDNAs to high levels and that the amount of fully assembled fibrinogen secreted is equal to the level of the lowest expressing chain.iii

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“…6, 23) interested in production of therapeutic human proteins in genetically engineered domestic animals have chosen the lactating mammary gland for transgene expression. Genes coding for therapeutic proteins have been successfully integrated and expressed in mammary glands of sheep (5, 23), goats (6), and pigs (21). Although milk production from these animals is copious, that of the dairy cow is superior; thus, the cow is an excellent choice for large-scale production of foreign proteins in the mammary gland.…”

Section: Introductionmentioning

confidence: 99%

Effects of Time of Deoxyribonucleic Acid Microinjection on Gene Detection and In Vitro Development of Bovine Embryos

Canseco

Russell

Johnson

et al. 1994

Journal of Dairy Science

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In vivo fertilized embryos were surgically collected from superovulated dairy cows to evaluate microinjection on embryo development and utilized the polymerase chain reaction technique for selection of transgenic embryos. Seventy-two percent of the embryos with visible pronuclei or nuclei were microinjected with DNA, and the remaining 28% served as uninjected controls. All embryos were cocultured with bovine oviductal epithelial cells. Mean final development scores of embryos within the same initial cell stage at collection were unaffected by microinjection. After 144 h of culture, 45% of the microinjected embryos developed to the morula or blastocyst stage. The transgene was detected in 50, 10, and 9% of demimorulae from embryos microinjected at the 1-, 2-, and 4-cell stages. Frequency of transgene detection was higher in morulae from 1-cell embryos than in morulae from 2-and 4-cell embryos. Use of in vitro coculture, embryo bisection, and polymerase chain reaction technique facilitated selection of bovine embryos that carried the transgene. (Key words: deoxyribonucleic acid microinjection, bovine embryo culture) Abbreviation key: OEC = oviductal epithelial cells, PCR = polymerase chain reaction.

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High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

Cited by 150 publications

References 17 publications

A perspective on fish gonad manipulation for biotechnical applications

A perspective on fish gonad manipulation for biotechnical applications

Production and secretion of recombinant human fibrinogen by the transgenic murine mammary gland

Effects of Time of Deoxyribonucleic Acid Microinjection on Gene Detection and In Vitro Development of Bovine Embryos

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