Generation of transgenic chickens that produce bioactive human granulocyte‐colony stimulating factor

Kwon, Mo Sun; Koo, Bon Chul; Choi, Bong Ryong; Park, Yoon-Yub; Lee, Young Man; Suh, Hun Suk; Park, Young Sik; Lee, Hoon Taek; Kim, Jin‐Hoi; Roh, Ji Yeol; Kim, Nam‐Hyung; Kim, Teoan

doi:10.1002/mrd.20860

Cited by 34 publications

(20 citation statements)

References 22 publications

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“…(9)(10)(11)(12) Biopharmaceuticals can thus be produced with high yield in egg white, which offers a sterile compartment with a stable and well-characterised protein composition. Several human recombinant proteins have been successfully produced in recent years, including human interferon a2b (13) and b1a, (14) and human granulocyte-colony stimulating factor, (15) with more expected to become available in coming years.…”

Section: Transgenic Chick Egg As Bioreactormentioning

confidence: 99%

The chick embryo: hatching a model for contemporary biomedical research

Rashidi

Sottile

2009

BioEssays

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Animal models play a crucial role in fundamental and medical research. Progress in the fields of drug discovery, regenerative medicine and cancer research among others are heavily dependent on in vivo models to validate in vitro observations, and develop new therapeutic approaches. However, conventional rodent and large animal experiments face ethical, practical and technical issues that limit their usage. The chick embryo represents an accessible and economical in vivo model, which has long been used in developmental biology, gene expression analysis and loss/gain of function experiments. It is also an established model for tissue/cell transplantation, and because of its lack of immune system in early development, the chick embryo is increasingly recognised as a model of choice for mammalian biology with new applications for stem cell and cancer research. Here, we review novel applications of the chick embryo model, and discuss future developments of this in vivo model for biomedical research.

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Section: Transgenic Chick Egg As Bioreactormentioning

confidence: 99%

The chick embryo: hatching a model for contemporary biomedical research

Rashidi

Sottile

2009

BioEssays

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“…Despite this success, long-term in vitro culture is generally held to be associated with reduced ability to colonize the gonad, although other researchers have successfully applied this technique (Park et al 2008, Shine et al 2009). …”

Section: Genetic Modification Of Pgcsmentioning

confidence: 99%

“…From 129 injected eggs, 13 hatched after 21 days of incubation; EGFP sequences were detected in several different tissues. More recently, Kwon (2008) used a similar technique to generate transgenic chickens producing recombinant human granulocyte colony-stimulating factor (hG-CSF); the biological activity of the recombinant protein was reported to be higher than that of hG-CSF produced commercially in E. coli.…”

Section: Introductionmentioning

confidence: 99%

Primordial germ cells (PGCs) as a tool for creating transgenic chickens

Chojnacka-Puchta¹,

Kasperczyk²,

Płucienniczak³

et al. 2012

Polish Journal of Veterinary Sciences

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The transgenic chicken has great potential as a bioreactor for the production of valuable pharmaceutical proteins, notably in the oviduct/egg. Whereas conventional transgenic approaches have significant limitations in this species, an alternative approach employing primordial germ cells (PGCs), the progenitor cells to ova and spermatozoa, has now been successfully applied to the insertion of exogenous genes into birds. Recent developments in manipulating avian embryos make it possible to produce germline chimeras derived from transferred PGCs. In this review we describe the migration pathway of chicken PGCs during early development. We then summarize different methods for the isolation of PGCs and the diversity of techniques used to introduce genes into these cells. Finally, we describe an in vitro assay for testing tissue-specific vectors designed to express heterologous proteins in transgenic chickens.

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“…The high efficacy of these vector systems for poultry transgenesis has been shown [1][2][3][4][5][6]. One of the main directions in the transgenesis of poultry is the development of effective vector systems, which provide the high expression of recombinant protein in the oviduct cells to produce a transgenic product.…”

Section: Introductionmentioning

confidence: 99%

Influence of Ovalbumin Gene Regulatory Elements on Tissue Specificity and Level of Transgene Expression

Волкова¹,

Fomin²,

Dotsev³

et al. 2016

S-h. biol.

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A b s t r a c tThe use of lentiviral vectors for the genetic modification of embryonic chicken cells is regarded as one of the promising methods for producing transgenic poultry. In this case, it is very important to determine the regulatory elements of the ovalbumin gene, providing tissue-specificity and high transgene expression in cells of chicken oviduct. The aim of this work was to study the effect of the intron sequences and promoter of ovalbumin gene on tissue specificity and level of the transgene expression. For this purpose constructs based on lentiviral vector pWpxl, containing eGFP marker gene under control of the modified ovalbumin gene regulatory elements were obtained. Vector pW2.8 included a chromosomal DNA fragment of 2.8 kb comprising a first exon, intron sequence and part of the second exon of the ovalbumin gene; vector pW1.2 -chromosomal DNA fragment of 1.2 kb comprising a promoter and ovalbumin gene sequence (without the intron and the first exon) to the transcription initiation point; vectors pW131, pW225, pW315 -chromosomal DNA fragment, similar to the fragment of 1.2 kb in pW1.2 vector in which promoter (80 bp) was replaced by highly structured sequence of the birds -actin gene promoter region of 131, 225 or 315 bp respectively. The control vector pWCAGgfp included constitutive hybrid regulatory element comprising human cytomegalovirus early gene enhancer and birds' -actin gene promoter (CAG). Primary culture cells of chick oviduct and human fibrosarcoma cells 293T (control) were used as target cells for transfection. Viral preparation was added after a monolayer of cells reached concentration of 1-3½10 7 CFU/ml. eGFP expression was determined by fluorimetry in 72 hours after transfection. Low level of expression of eGFP gene controlled by chromosomal fragment of 2.8 kb leader region of the ovalbumin gene was confirmed in vitro using culture of chicken oviduct cells and 293T cell line: in vector pW2.8 recombinant protein expression level was up to 25 times lower compared to pWCAGgfp vector with a constitutive promoter CAG. Yet the eGFP expression levels for pW1.2 and pW2.8 constructs were identical, indicating the absence of introns' influence on the expression level of the recombinant DNA using this regulatory element. When the ovalbumin gene promoter was replaced by highly structured elements of -actin constitutive gene promoter the increase in the expression of eGFP in 2-3 times was observed, as well as the increased expression occurred with lengthening of the promoter region of -actin gene (vectors pW131, pW225, pW315). The achieved levels of expression with the use of exogenous -actin gene promoter were comparable with expression levels controlled by a constitutive promoter CAG, however when the promoter part of the ovalbumin gene was replaced by exogenous promoter (gene -actin), the deregulation of tissue-specific expression of eGFP was observed, indicating that transcription with tissue-specific ovalbumin promoter gene can be modulated or activated with exogenous enhancers.

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Generation of transgenic chickens that produce bioactive human granulocyte‐colony stimulating factor

Cited by 34 publications

References 22 publications

The chick embryo: hatching a model for contemporary biomedical research

The chick embryo: hatching a model for contemporary biomedical research

Primordial germ cells (PGCs) as a tool for creating transgenic chickens

Influence of Ovalbumin Gene Regulatory Elements on Tissue Specificity and Level of Transgene Expression

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