Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation
Joint Authors
Hwang, In-Sul
Kwon, Dae-Jin
Lee, Jeong-Woong
Hwang, Seongsoo
Sun, Wu-Sheng
Chun, Ju-Lan
Do, Jeong-Tae
Kim, Dong-Hwan
Ahn, Jin-Seop
Kim, Min-Kyu
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2015), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-11-24
Country of Publication
Egypt
No. of Pages
11
Abstract EN
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 naïve 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 naïve pluripotent stem cells from other metastable state cells in a heterogenous population.
American Psychological Association (APA)
Sun, Wu-Sheng& Chun, Ju-Lan& Do, Jeong-Tae& Kim, Dong-Hwan& Ahn, Jin-Seop& Kim, Min-Kyu…[et al.]. 2016. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation. Stem Cells International،Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1116029
Modern Language Association (MLA)
Sun, Wu-Sheng…[et al.]. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation. Stem Cells International Vol. 2016, no. 2016 (2015), pp.1-11.
https://search.emarefa.net/detail/BIM-1116029
American Medical Association (AMA)
Sun, Wu-Sheng& Chun, Ju-Lan& Do, Jeong-Tae& Kim, Dong-Hwan& Ahn, Jin-Seop& Kim, Min-Kyu…[et al.]. Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation. Stem Cells International. 2016. Vol. 2016, no. 2016, pp.1-11.
https://search.emarefa.net/detail/BIM-1116029
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1116029