Deposition of bioactive human epidermal growth factor in the egg white of transgenic hens using an oviduct‐specific minisynthetic promoter

Abstract

Currently, transgenic animals have found a wide range of industrial applications and are invaluable in various fields of basic research. Notably, deposition of transgene-encoded proteins in the egg white (EW) of hens affords optimal production of genetically engineered biomaterials. In the present study, we developed a minisynthetic promoter modulating transgene transcription specifically in the hen's oviduct, and assayed the bioactivity of human epidermal growth factor (hEGF) driven by that promoter, after partial purification of epidermal growth factor (EGF) from transgenic hen eggs. Our minisynthetic promoter driving expression of chicken codon-optimized human epidermal growth factor (cEGF) features 2 consecutive estrogen response elements of the ovalbumin (OV) promoter, ligated with a 3.0 kb OV promoter region carrying OV regulatory elements, and a 59-UTR. Subsequently, a 39-UTR carrying the poly-A tail sequence of the OV gene was added after incorporation of the cEGF transgene. Finally, we partially purified cEGF from transgenic hen eggs and evaluated the biofunctional activities thereof in vitro and in vivo. In the in vitro assay, EW-derived hEGF exhibited a proliferative effect on HeLa cells similar to that of commercial hEGF. In the in vivo assay, compared to the nontreated control, transgenic hen egg-derived EGF afforded slightly higher levels of re-epithelialization (via fibroplasia) and neovascularization of wounded skin of miniature pigs than did the commercial material. In conclusion, transgenic hens may be used to produce genetically engineered bioactive biomaterials driven by an oviduct-specific minisynthetic promoter.-Park, T.

Three-dimensional culture of chicken primordial germ cells in chemically defined media containing the functional polymer FP003

“…Eggs are considered an ideal platform for recombinant protein production, and the ovalbumin promoter shows a robust and specific expression ability in oviducts. Therefore, numerous studies have attempted to produce recombinant proteins using transgenic hens [13, 16, 17]. In addition, oviduct bioreactor also presents the easiness in transgenic animal production and husbandry, as well as the recombinant protein purification.…”

“…Transgenic hen as bioreactor to produce therapeutic protein in laid egg has been produced and some of the lines show high to low productivity of the proteins of interest [16, 17]. However, it is tedious to generate each transgenic chicken line and to select strains with the better productive efficiency.…”

CRISPR/Cas9 gene editing in a chicken model: current approaches and applications

“…The use of chickens as bioreactors provides an alternative therapeutic approach, which greatly benefits human health (Zhu et al 2005). Previous studies demonstrated using modified chickens to produce, for example, antimicrobial peptides , monoclonal antibodies for breast cancer therapy (Oishi et al 2011), lysozyme , epidermal growth factor (Park et al 2015), and human cytokine interferon α2a (Herron et al 2018). Recently, Oishi et al (2018) has proven that the CRISPR/Cas9 system can be successfully used to produce recombinant proteins in egg whites.…”

SC1 sustains the self‐renewal capacity and pluripotency of chicken blastodermal cells by inhibiting the phosphorylation of ERK1 and promoting the phosphorylation of Akt

“…They are also used as an ideal substitute for necrosed tissue to treat diseases, including degenerative diseases (Mead et al, 2015). Transgenic chicken embryo is one of the most effective and exciting research tools in the field of agriculture and medicine (Cao et al, 2015;Li et al, 2014;Park et al, 2015;Zhu et al, 2005). Furthermore, chicken blastodermal cells (cBCs) can be utilized as a model to investigate self-renewal, pluripotency and epigenesis of ES cells (Busch, Krochmann, & Drews, 2013;Cloney & Franz-Odendaal, 2015;Jahanpanah et al, 2014;Kress, Montillet, Jean, Fuet, & Pain, 2016;Li et al, 2014).…”