Earticle

초록

영어

Porcine spermatogonial stem cells (SSCs) are an useful model for a successful gene modification of sperm compatible, and various gene delivery systems to transport specific genes into the cytoplasm of porcine SSCs have been introduced for successfully producing transgenic sperms. However, each gene delivery system still suffers from low transfection efficiency and transfected gene expression in porcine SSCs, resulting in making it difficult to produce transgenic offspring. Accordingly, in order to determine type of promoters optimized to porince SSCs, impact of each Cytomegalovirus (CMV), Simian virus 40 (SV40), and chicken b-actin promoter on transfection efficiency of porcine SSCs was investigated using electroporation. For these, the highest transfection efficiency and cell viability were observed in porcine SSCs transfected with 1 μg of the each transgenic vector with a single electric pulse from an electroporator at a voltage of 200 V and a capacitor setting of 500 μF in a 0.4 cm cuvette containing 200μL of standized SSCs medium. Cell viability was measured by trypan blue exclusion assay, and analysis of transfection efficiency and intensity were conducted as counting SSCs expressing enhanced green fluorescent protein (EGFP) and measuring EGFP fluorescent intensity under flow cytometer system. As the results, the highest EGFP expression efficiency and intensity were observed in porcine SSCs introduced with CMV promoter-containing vectors, compared to those with SV40 and chicken b-actin protmoter- containing vectors. However, cell viability showed no significant difference among porcine SSCs experiencing transfection of SV40, CMV and chicken b-actin promotercontaining vector. In conclusion, we found that CMV promoter could effectively stimulate expression of genes transfected to porcine SSCs. This will contribute advances in further research related in SSCs transplantation and production of transgenic animals.

키워드

저자

• Min Seong Kim [ Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Min Hee Park [ Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Ji Eun Park [ Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Seong Jae Kim [ Department of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Jung Im Yun [ Division of Animal Resource Science Kangwon National University, Chuncheon 24341, Korea ]
• Eunsong Lee [ College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea ]
• Seung Tae Lee [ Department of Animal Life Science, College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  발생공학 국제심포지엄 및 학술대회 [International Symposium on Developmental Biotechnology]

• 간기

  연간

• 수록기간

  2004~2018

• 십진분류

  KDC 527 DDC 636