Construction of a dual-fluorescence reporter system to monitor the dynamic progression of pluripotent cell differentiation

Cited 3 time in scopus
Metadata Downloads
Title
Construction of a dual-fluorescence reporter system to monitor the dynamic progression of pluripotent cell differentiation
Author(s)
W S Sun; J L Chun; J T Do; Dong Hwan Kim; Jin Seop Ahn; M K Kim; I S Hwang; D J Kwon; S S Hwang; Jeong Woong Lee
Bibliographic Citation
Stem Cells International, vol. 2016, pp. 1390284-1390284
Publication Year
2016
Abstract
Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE) and proximal enhancer (PE), in the 5′ upstream regulatory sequences (URSs) of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs). We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free-Oct4-promoter-driven EGFP reporter cassette with a PE-free-Oct4-promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9) and a mouse EpiSC-like cell line (P19) before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naive pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naive pluripotent stem cells from other metastable state cells in a heterogenous population.
ISSN
1687-966X
Publisher
Hindawi Ltd
DOI
http://dx.doi.org/10.1155/2016/1390284
Type
Article
Appears in Collections:
Division of Biomedical Research > Biotherapeutics Translational Research Center > 1. Journal Articles
Files in This Item:

Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.