In vivo and in vitro expression of human serum albumin genomic sequences in mammary epithelial cells with ?-lactoglobulin and whey acidic protein promoters

Barash, Itamar; Faerman, Alexander; Richenstein, Moshe; Kari, Rachel; Damary, Golda-Meir; Shani, Moshe; Bissell, Mina J.

doi:10.1002/(sici)1098-2795(199903)52:3<241::aid-mrd1>3.0.co;2-x

Cited by 20 publications

(11 citation statements)

References 49 publications

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“…We assume that the fragment of WAP promoter used in our study included essential sequences required for directing expression exclusively to the mammary gland, since ectopic expression of the rhGH gene construct was not observed in this study, in spite of observations concerning the ectopic expression of other proteins under the WAP promoter (Wall et al 1995; Barash et al 1999). …”

Section: Discussionmentioning

confidence: 94%

Expression of human growth hormone in the milk of transgenic rabbits with transgene mapped to the telomere region of chromosome 7q

et al. 2012

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The advent of transgenic technology has provided methods for the production of pharmaceuticals by the isolation of these proteins from transgenic animals. The mammary gland has been focused on as a bioreactor, since milk is easily collected from lactating animals and protein production can be expressed at very high levels, including hormones and enzymes. We demonstrate here the expression pattern of recombinant human growth hormone (rhGH) in transgenic rabbits carrying hGH genomic sequences driven by the rat whey acidic protein (WAP) promoter. The transgene was mapped to the q26-27 telomere region of chromosome 7q by fluorescence in situ hybridization (FISH). Nearly 30 % of the F1 generation demonstrated the presence of transgene. The recombinant growth hormone was detected in the milk of the transgenic rabbit females, but not in serum, up to the level of 10 μg/ml. Ectopic expression of the transgene in the brain, heart, kidney, liver, and salivary gland was not observed, indicating that a short sequence of rat WAP promoter (969 bp) contained essential sequences directing expression exclusively to the mammary gland. The biological activity of recombinant growth hormone was measured by immunoreactivity and the capability to stimulate growth of the hormone-dependent Nb211 cell line.

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Section: Discussionmentioning

confidence: 94%

Expression of human growth hormone in the milk of transgenic rabbits with transgene mapped to the telomere region of chromosome 7q

et al. 2012

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“…Another possibility may be a greater efficiency of the WAP and as 1 -casein constructs in driving IGF-I over-expression than the a-LA promoter. To our knowledge, direct comparisons between the efficiency of a-LA and other milk protein promoters has not been conducted, however, the WAP promoter was shown to be less efficient than the b-lactoglobulin promoter at driving the over-expression of human serum albumin into milk (Barash et al, 1999). The highest level achieved by the WAP promoter was an order of magnitude lower than the b-lactoglobulin promoter (Barash et al, 1999).…”

Section: Discussionmentioning

confidence: 95%

Mammary Specific Transgenic Over-expression of Insulin-like Growth Factor-I (IGF-I) Increases Pig Milk IGF-I and IGF Binding Proteins, with no Effect on Milk Composition or Yield

et al. 2005

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IGF-I regulates lactation by stimulating mammary mitogenesis, inhibiting apoptosis, and partially mediating the effects of growth hormone on lactogenesis. Herein, lactation performance during first and second parity was assessed in transgenic swine (TG) that over-expressed human IGF-I in milk under the control of the bovine alpha-lactalbumin promoter, regulatory regions and signal peptide coding sequence. Milk samples were collected throughout lactation (farrowing to d24) from TG sows and non-transgenic littermates (CON) and IGF-I, IGF-II, and IGFBP determined. Colostral (<24 h postpartum) IGF-I content was 26-fold greater (p<0.001) in TG sows (949+/- 107 microg/L; range 228-1,600 microg/L) than CON (36+/-17.8 microg/L) and was 50- to 90-fold greater (p< 0.001) in mature milk (d2-24 postpartum). There was no effect of parity on milk IGF-I content. Milk IGF-II concentration was unaffected by IGF-I over-expression. Low molecular weight IGFBP (IGFBP-2 and -5) in the milk of TG sows were higher (p=0.02) than CON in the early postpartum period, but did not differ in mature milk. Milk yield, determined by weigh-suckle-weigh, was similar in TG and CON as was litter weight gain. Milk nutrient composition was not significantly affected by IGF over-expression. Thus, mammary specific transgenic over-expression of IGF-I significantly increased milk IGF-I and IGFBP content, but did not impact lactation performance in swine.

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“…(18) The WAP promoter was shown to be less efficient than the b-lactoglobulin promoter at driving the overexpression of recombinant human protein into milk. (19) However, in our transgenic mice we confirmed high levels of tissue-specific expression of hFVIII in the mammary glands. On the other hand, hFVIII has been produced in the milk of transgenic animals.…”

Section: Figmentioning

confidence: 51%

Investigation of hFVIII Production in Mammary Glands of Transgenic Mice

Mohammadian

Rassi²

2014

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Hemophilia A is an X-linked disorder affecting 1 in 10,000 males. The disease is caused by a defect or mutation of factor 8 or 9. Human factor 8 gene (hFVIII) is a relatively large gene consisting of 26 exons and approximately 2,351 amino acids with a length of 9 Kb mRNA. Expression of hFVIII in mammalian milk is becoming a widespread strategy for high-level production of hFVIII because of the most complex post-translational modifications. The aim of this study was the cloning and expression of hFVIII in mammary glands of two transgenic mice. To obtain a recombinant plasmid, first a plasmid carrying an FVIII gene fragment (pCMV6-hFVIII) was digested by EcoRI-SalI restriction enzymes and then the fragment was purified from agarose gel and inserted into a pUCWAP7 vector carrying a tissue-specific promoter (mWAP 4.1 kbp). After that, it was isolated by agarose gel and transferred into the murine zygotes by standard microinjection methods. Methods for expression of recombinant FVIII RT-PCR and ELISA were studied. The results show the successful expression of factor FVIII gene and its product in the mouse mammary glands.

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In vivo and in vitro expression of human serum albumin genomic sequences in mammary epithelial cells with ?-lactoglobulin and whey acidic protein promoters

Cited by 20 publications

References 49 publications

Expression of human growth hormone in the milk of transgenic rabbits with transgene mapped to the telomere region of chromosome 7q

Expression of human growth hormone in the milk of transgenic rabbits with transgene mapped to the telomere region of chromosome 7q

Mammary Specific Transgenic Over-expression of Insulin-like Growth Factor-I (IGF-I) Increases Pig Milk IGF-I and IGF Binding Proteins, with no Effect on Milk Composition or Yield

Investigation of hFVIII Production in Mammary Glands of Transgenic Mice

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