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KMID : 1005220150300030219
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Journal of Embryo Transfer
2015 Volume.30 No. 3 p.219 ~ p.224
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Production of hGCSF and GFP Co-Expressed Transgenic Cow Embryo by Somatic Cell Nuclear Transfer Technique
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Yang Jung-Seok
Joe So-Young
Koo Bon-Chul
Heo Young-Tae
Lee Su-Min
Kang Man-Jong
Song Hyuk
Ko Dae-Hwan
Uhm Sang-Jun
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Abstract
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The purpose of this study is to develop transgenic cell line expressing targeted human granulocyte colony stimulating factor (hGCSF) and green fluorescence protein (GFP) genes as well as production of Somatic Cell Nuclear Transfer (SCNT) embryos derived from co-expressed transgenic donor cells. Constructed pPiggy-mWAP-hGCSF-EF1-GFP vector was chemically transfected into bovine fetus cells and then, only GFP expressed cells were selected as donor cells for SCNT. Cleavage and blastocyst rates of parthenogenetic, SCNT embryos using non-TG cell and hGCSF-GFP dual expressed SCNT embryos were examined (cleavage rate: 78.0±2.8 vs. 73.1±3.2 vs. 70.4±4.3%, developmental rate: 27.2 ±3.2 vs. 21.9±3.1 vs. 17.0±2.9%). Result indicated that cleavage and blastocyst rates of TG embryos were significantly lower (P<0.05) than those of parthenogenetic and non-TG embryos, respectively. In this study, we successfully produced hGCSF-GFP dual expressed SCNT embryos and cryopreserved to produce transgenic cattle for bioreactor system purpose. Further process of our research will transfer of transgenic embryos to recipients and production of hGCSF secreting cattle.
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KEYWORD
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SCNT , hGCSF , GFP , transgenic embryo , cow
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