원문정보
초록
영어
The transcription factor OCT4 has a critical role in central regulator of maintaining pluripotency and self-renewal. Also, it is an important marker of undifferentiating status in early mammalian embryonic development and embryonic stem cells. There are four conserved regions (CR1, CR2, CR3 and CR4) in the 5’ upstream regulatory region of various species. It contains core promoter and two conserved enhancers, distal enhancer (DE) and proximal enhancer (PE). Even though study related dual-fluorescence reporter was reported, there is still a dearth of sufficient information about porcine OCT4 upstream reporter system until now. So, the purpose of this study is investigation to find conserved regions among other species using comparative sequence analysis of porcine OCT4, and conduct functional analysis of porcine OCT4 promoter region. And we designed four constructs containing the different elements of the OCT4 promoter and enhancers in luciferase vectors, and they were transfected into mouse embryonic stem cells and embryonic carcinoma cells. Our results indicate that porcine OCT4 promoter and regulatory regions have a high similarity with other mammalian species. Four porcine OCT4-luciferase constructs showed different expression patterns in mouse embryonic stem cells, mouse embryonic carcinoma cells, and mouse embryonic fibroblast. And we designed four constructs containing the different elements of the OCT4 promoter and enhancers in GFP, RFP vectors and they were transfected into mouse embryonic stem cells. Those constructs can work in mouse embryonic stem cells and mouse embryonic carcinoma cells. This work could provide basic information for porcine OCT4 upstream region and suggest the various type of porcine OCT4-Fluoresence reporter constructs, which can use for reporter system of studying species-specific pluripotency in early embryo developments and establishment of embryonic stem cells.