Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
권호 표지
DOI QR코드

DOI QR Code

Production of hGCSF and GFP Co-Expressed Transgenic Cow Embryo by Somatic Cell Nuclear Transfer Technique

체세포 핵치환 기술을 이용한 hGCSF와 GFP 유전자 동시발현 형질전환 소 배아 생산

  • Yang, Jung Seok (Dept. of Animal Science and Biotechnology, Sangji Youngseo College) ;
  • Joe, So Young (Dept. of Animal Science and Biotechnology, Sangji Youngseo College) ;
  • Koo, Bon-Chul (Dept. of Physiology, Catholic University of Daegu School of Medicine) ;
  • Heo, Young-Tae (Dept. of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Lee, Su Min (Dept. of Animal Science and Biotechnology, Sangji Youngseo College) ;
  • Kang, Man-Jong (Dept. of Animal Science, Chunnam National University) ;
  • Song, Hyuk (Dept. of Animal Biotechnology, College of Animal Bioscience & Technology, Konkuk University) ;
  • Ko, Dae Hwan (Dept. of Animal Science and Biotechnology, Sangji Youngseo College) ;
  • Uhm, Sang Jun (Dept. of Animal Science and Biotechnology, Sangji Youngseo College)
  • 양정석 (상지영서대학교 동물생명산업과) ;
  • 조소영 (상지영서대학교 동물생명산업과) ;
  • 구본철 (대구가톨릭대학교 의학과) ;
  • 허영태 (건국대학교 동물생명공학과) ;
  • 이수민 (상지영서대학교 동물생명산업과) ;
  • 강만종 (전남대학교 축산학과) ;
  • 송혁 (건국대학교 동물생명공학과) ;
  • 고대환 (상지영서대학교 동물생명산업과) ;
  • 엄상준 (상지영서대학교 동물생명산업과)
  • Received : 2015.06.05
  • Accepted : 2015.08.24
  • Published : 2015.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

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: $78.0{\pm}2.8$ vs. $73.1{\pm}3.2$ vs. $70.4{\pm}4.3%$, developmental rate: $27.2{\pm}3.2$ vs. $21.9{\pm}3.1$ vs. $17.0{\pm}2.9%$). 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.

Keywords

References

  1. Batista RI, Melo CH, Souza-Fabjan JM, Teixeira DI, Melo LM and Freitas VJ. 2014. Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk. Biotechnol. Lett. 36:2155-2162. https://doi.org/10.1007/s10529-014-1588-0
  2. Cho SK, Kim JH, Park JY, Choi YJ, Bang JI, Hwang KC, Cho EJ, Sohn SH, Uhm SJ, Koo DB, Lee KK, Kim T and Kim JH. 2007. Serial cloning of pigs by somatic cell nuclear transfer: Restoration of phenotypic normality during serial cloning. Dev. Dyn. 236:3369-3382. https://doi.org/10.1002/dvdy.21308
  3. Choe CY, Son JK, Cho SR, Kang DW, Yeon SH, Choi SH, Choi SH, Kim NT, Kim JB, Jung YS, Kim SJ, Jung JW, Bok NH, Yoo YH and Son DS. 2010. Effects of pregnant rate after embryo transfer in oxygen consumption of embryos in Korean cattle. Korean J. Emb. Trans. 25:145-148.
  4. Choe CY, Son JK, Cho SR, Kang DW, Yeon SH, Choi SH, Kim NT, Jung YS, Kim SJ, Jung JW, Bok NH, Choi JS and Son DS. 2011. Effects of corpus luteum grade of estrus synchronized recipients on pregnancy rate following embryo transfer in Korean cattle. Korean J. Emb. Trans. 26:33-36.
  5. Doll DC and Weiss RB. 1983. Chemotherapeutic agents and the erythron. Cancer Treat. Rev. 103:185-200.
  6. Hofmann A, Kessler B, Ewerling S, Weppert M, Vogg B, Ludwig H, Stojkovic M, Boelhauve M, Brem G, Wolf E and Pfeifer A. 2003. Efficient transgenesis in farmanimals by lentiviral vectors. EMBO Rep. 4:1054-1060. https://doi.org/10.1038/sj.embor.7400007
  7. Hofmann A, Zakhartchenko V, Weppert M, Sebald H, Wenigerkind H, Brem G, Wolf E and Pfeifer A. 2004 Generation of transgenic cattle by lentiviral gene transfer into oocytes. Biol. Reprod. 71:405-409. https://doi.org/10.1095/biolreprod.104.028472
  8. Jaenisch R. 1976. Germ line integration and Mendelian transmission of the exogenous Moloney leukemia virus. Proc. Natl. Acad. Sci. USA 73:1260-1264. https://doi.org/10.1073/pnas.73.4.1260
  9. Kim YJ, Park H, Lee HL, Shin DS, Jo SW, Kim YS and Kim SH. 2008. Results of embryo transfer with Hanwoo embryos produced in-vivo or in-vitro to Holstein cows as recipients. Korean J. Emb. Trans. 23:167-175.
  10. Koo BC, Kwon MS, Roh JY, Kim M, Kim JH and Kim T. 2012. Quantitative analysis of tetracycline-inducible expression of the green fluorescent protein gene in transgenic chickens. J. Reprod. Dev. 58:672-677. https://doi.org/10.1262/jrd.2012-092
  11. Krestel HE, Mihaljevic AL, Hoffman DA and Schneider A. 2004. Neuronal co-expression of EGFP and beta-galactosidase in mice causes neuropathology and premature death. Neurobiol. Dis. 17:310-318. https://doi.org/10.1016/j.nbd.2004.05.012
  12. Lee GS, Jeong YW, Kim JJ, Park SW, Ko KH, Kang M, Kim YK, Jung EM, Moon C, Hyun SH, Hwang KC, Kim NH, Shin T, Jeung EB and Hwang WS. 2014. A canine model of Alzheimer's disease generated by overexpressing a mutated human amyloid precursor protein. Int. J. Mol. Med. 33: 1003-1012. https://doi.org/10.3892/ijmm.2014.1636
  13. Mastromonaco GF and King WA. 2007. Cloning in companion animal, non-domestic and endangered species: Can the technology become a practical reality? Reprod. Fertil. Dev. 19:748-761. https://doi.org/10.1071/RD07034
  14. Miao X. 2013. Recent advances in the development of new transgenic animal technology. Cell. Mol. Life Sci. 70:815-828. https://doi.org/10.1007/s00018-012-1081-7
  15. Rideout WM, Eggan K and Jaenisclh R. 2001. Nuclear cloning and epigenetic reprogramming of the genome. Science. 293: 1093-1098. https://doi.org/10.1126/science.1063206
  16. Samoilov AV, Kesian AZ and Suraeva NM. 2013. Development of transgenic chicken with a gene of human granulocyte colony-stimulating factor using sperm-mediated gene transfer. Izv. Akad. Nauk. Ser. Biol. 5:517-521.
  17. Steinberg SE, Campbell CL, Rabinovitch PS and Hillman RS. 1980. The effect of trimethoprim/sulfamethoxazole on friend erythroleukemia cells. Blood. 1980 553:501-504.
  18. Uhm SJ, Kim NH, Kim T, Chung HM, Chung KH, Lee HT and Chung KS. 2000. Expression of enhanced green fluorescent protein (EGFP) and neomycin resistant (Neo(R) genes in porcine embryos following nuclear transfer with porcine fetal fibroblasts transfected by retrovirus vector. Mol. Reprod. Dev. 57:331-337. https://doi.org/10.1002/1098-2795(200012)57:4<331::AID-MRD4>3.0.CO;2-7
  19. Xu YN, Uhm SJ, Koo BC, Kwon MS, Roh JY, Yang JS, Choi HY, Heo YT, Cui XS, Yoon JH, Ko DH, Kim T and Kim NH. 2013. Production of transgenic Korean native cattle expressing enhanced green fluorescent protein using a FIVbased lentiviral vector injected into MII oocytes. J. Genet. Genomics 40:37-43. https://doi.org/10.1016/j.jgg.2012.11.001