Production of Biofunctional Recombinant Human Interleukin 1 Receptor Antagonist (rhIL1RN) from Transgenic Quail Egg White1

Kwon, Soyoung; Choi, Jin Won; Jang, Hyun‐Jun; Shin, Sang Su; Lee, Seul Ki; Park, Tae Sub; Choi, In Young; Lee, Gwan Sun; Song, Gwonhwa; Han, Jae Yong

doi:10.1095/biolreprod.109.081687

Cited by 43 publications

(36 citation statements)

References 31 publications

(42 reference statements)

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“…Using transgenic poultry as bioreactors is one of the alternative methods of producing recombinant proteins [1][2][3][4][5][6][7][8][9]. Poultry is immune to potential therapeutic proteins (e.g., human erythropoietin), the expression of which can adversely affect the health of transgenic mammals when used as a commercial producers of drugs.…”

Section: Introductionmentioning

confidence: 99%

Efficiency of Local Transgenesis of the Oviductal Cells in Chicken as Influenced by Hormonal Stimulation

Белоглазов¹,

Волкова²,

Волкова³

et al. 2015

S-h. biol.

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A b s t r a c tOne of the promising areas of biotechnology is to create transgenic chicken bioreactors. However, despite notable successes in avian transgenesis, the creation of transgenic chickens now is a particular problem. Searches and development of alternative methods of directed gene transfer are required, one of which is genetic transformation of certain organs, in particular chicken oviduct (somatic transgenesis). This can significantly reduce the expenditure of time and materials in the preparation of transgenic organisms than using other cellular targets for targeted delivery of DNA (cell blastoderm, primordial germ cells, embryonic stem cells) because the genetically engineering manipulation is possible to be conducted only on embryonic material. The aim of this study was to investigate the effectiveness of the delivery of recombinant DNA into the chicken oviduct cells in vivo and development of methodological approaches to increase the efficiency of transgenesis. In the present work a retroviral vector pLN-GFP was used, based on the Moloney murine leukemia virus (Mo-MuLV), in which a sequence of GFP (green fluorescent protein) marker gene has been cloned. Packaging cell line GP+envAM12 has been used to package retroviral vector pLN-GFP. To determine the optimal duration of administration of gene constructs a proliferative activity was analyzed in the oviduct cells at the age of 1, 2, 3, 4, 4.5 and 5 months, and at the age of 2 months within 24 and 48 hours after hormonal stimulation by 0.1 % sinestrol solution. The introduction of the retroviral vector was performed by injecting the solution of the gene construct directly into the protein part of the oviduct of hens at the age of 4 months without hormonal pre-stimulation (group I) and at the age of 2 months within 24 h after sinestrol injection (group II). Analysis of the integration and expression of recombinant DNA in chick oviduct cells was performed at the age of 6 months. As a result of histological investigations it was found that the maximum proliferative activity in the protein part of oviduct (magnum) was observed in a period between 4 and 4.5 months: the relative DNA content of the cells at this period increased by 3.4 rel. units, which was equivalent to an increase of this parameter over the entire preceding period from 1 to 4 months. The analysis of histological sections of the oviduct in 2-month-old chickens 24 hours after sinestrol injection revealed a significant change in the structure of the oviduct, typical for histological structure of the magnum portion of the oviduct at the 4 month age, and after 48 hours the oviduct histological structure corresponded to that of mature chicken. An average efficiency of the hen oviduct cell transformation assessed as a percentage of the transformed cells to the total cell number of this type in the oviduct, in group I was 17.2±3.1 % whereas in group II it was 57.3±6.3 %. Thus, the use of hormone treatment has allowed a 3.3-fold increase in the effectiveness of local transgenesis of the oviductal...

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Section: Introductionmentioning

confidence: 99%

Efficiency of Local Transgenesis of the Oviductal Cells in Chicken as Influenced by Hormonal Stimulation

Белоглазов¹,

Волкова²,

Волкова³

et al. 2015

S-h. biol.

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“…By injecting a replication-defective pantropic retrovirus into quail blastoderms, Mizuarai et al (2001) reported the production of transgenic quail through germline transmission. Thereafter, the virus-mediated transgenic technique at stage X blastoderm demonstrated that transgenic quail expressed a biofunctional protein on the oviduct-specific manner and was used for studying vascular development and morphogenesis as an avian model (Kwon et al, 2010;Sato et al, 2010). The lentiviral transduced PGC-mediated transgenesis successfully produced transgenic quail although the efficiency was still too low (Shin et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Non-Viral Transgenesis via Direct In Ovo Lipofection in Quail

Park

Han²

2015

Korean Journal of Poultry Science

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Transgenic animals have been widely used for developmental biology studies, as disease models, and even in industry such as transgenic bioreactor animals. For transgenic birds, quail has the great advantages of small body size, short generation time, and frequent egg production. To date, retroviral or lentiviral transduction has been used to generate transgenic quail for various purposes. However, the efficiency of transgenic offspring production with these methods is relatively low and viral vector usage has safety issues. Unfortunately, non-viral transgenesis has not been established in quail due to a deficiency of stem cell and germ cell culture systems. In this study, we established a direct in ovo lipofection method that could be used to create transgenic quail without germline-competent cells or viruses. To optimize the injection stage during embryo development, the liposome complex (containing piggyBacCMV-GFP and transposase plasmids) was introduced into an embryonic blood vessel at 50 hr, 55 hr or 60 hr. GFP expression was detected in various tissues (heart, kidney, liver and stomach) on day 12 of incubation under a fluorescence microscope. Additionally, GFP-positive cells were detected in the recipient embryonic gonads. In conclusion, the direct in ovo lipofection method with the piggyBac transposon could be an efficient and useful tool for generating transgenic quail.

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“…Moreover, transgenic technology provides the possibility of utilizing egg-laying avian species as a production platform for the large-scale synthesis of target proteins. Oviduct-specific transcription elements, particularly the OVA promoter, are interesting tools that may be used in the production of specific target proteins (Kwon et al, 2010). Avian oviduct bioreactors have been widely studied in biotechnology, in which foreign proteins were found to be mainly expressed in the oviduct (Lillico et al, 2007;Kwon et al, 2010;Byun et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Molecular cloning, expression, and regulation of the ovalbumin gene in pigeon oviduct epithelial cells

et al. 2014

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ABSTRACT. The full-length pigeon ovalbumin (OVA) gene cDNA was cloned and sequenced by reverse transcription-polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends. A 386-amino acid protein was predicted for the obtained sequence, which had 67% identity with the chicken protein. Similar to chicken OVA, the pigeon OVA gene is a non-inhibitory serine protease inhibitor. Quantitative PCR analysis revealed that pigeon OVA mRNA was highly expressed in the oviduct, and trace amounts were detected in other tissues. During the reproductive cycle, pigeon oviduct OVA mRNA expression reached its peak during the egg-laying stage, decreased with brooding, and then increased again during the squabfeeding period. Moreover, the relative OVA expression level in pigeon oviduct epithelial cells could be upregulated by a constant concentration of steroid hormones.

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Production of Biofunctional Recombinant Human Interleukin 1 Receptor Antagonist (rhIL1RN) from Transgenic Quail Egg White1

Cited by 43 publications

References 31 publications

Efficiency of Local Transgenesis of the Oviductal Cells in Chicken as Influenced by Hormonal Stimulation

Efficiency of Local Transgenesis of the Oviductal Cells in Chicken as Influenced by Hormonal Stimulation

Non-Viral Transgenesis via Direct In Ovo Lipofection in Quail

Molecular cloning, expression, and regulation of the ovalbumin gene in pigeon oviduct epithelial cells

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