Establishment of a Porcine Oct-4 Promoter-Driven EGFP Reporter System for Monitoring Pluripotency of Porcine Stem Cells
Porcine pluripotent cells with the capacity to generate germ line chimeras have not been developed yet. The transcription factor Oct-4 is an important marker of undifferentiating status and a central regulator of pluripotency in cells. Establishment of an Oct-4 promoter-based reporter system, such as that used in mice, will be a useful tool for monitoring the differentiating statuses of porcine cells both in vivo and in vitro. In the present study, we constructed a vector, pOGN2, in which enhanced green fluorescent protein (EGFP) was driven by the porcine Oct-4 promoter. In pigs containing this vector, EGFP was expected to be specifically expressed in pluripotent cells. We delivered the vectors into porcine fetal fibroblasts (PEFs) using liposomes. After transfected PEFs were selected with G418, we established eight cell lines containing the pOGN2 vector. When transgenic cells were used as donor nuclei to make somatic cell nuclear transfer (SCNT) embryos, SCNT embryos derived from four transgenic cell lines expressed green fluorescence. When PEFs with pOGN2 vectors were infected with retroviral vectors encoding the four transcription factors (Oct-4, Sox2, Klf4, and c-Myc), EGFP-expressing iPS cell colonies were observed at day 20. This work lays a foundation that can be used to generate a pig strain with an Oct4-EGFP reporter system, which would be greatly helpful in studying the differentiating and reprogramming mechanisms of pig embryos.
Lizhen Huang Nana Fan Jie Cai Dongshan Yang Bentian Zhao Zhen Ouyang Weiwang Gu Liangxue Lai
Institute of Comparative Medicine and Center of Experimental Animals Southern Medical University,Gua Key Laboratory of Regenerative Biology ,South Chian Institue for Stem Cell Biology and Regenerative Key Laboratory of Regenerative Biology ,South Chian Institue for Stem Cell Biology and Regenerative
国内会议
2011东莞第二届国际小型猪学术论坛暨大型实验动物生物医药研究应用研讨会
东莞
英文
285-290
1900-01-01（万方平台首次上网日期，不代表论文的发表时间）