Construction and Quantitative Evaluation of a Dual Specific Promoter System for Monitoring the Expression Status of Stra8 and c-kit Genes.

Details

Serval ID
serval:BIB_8E71E0F4AAF1
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Construction and Quantitative Evaluation of a Dual Specific Promoter System for Monitoring the Expression Status of Stra8 and c-kit Genes.
Journal
Molecular Biotechnology
Author(s)
Dastpak M., Matin M.M., Farshchian M., Arsenijevic Y., Momeni-Moghaddam M., Sisakhtnezhad S., Boozarpour S., Bidkhori H.R., Mirahmadi M., Bahrami A.R.
ISSN
1559-0305 (Electronic)
ISSN-L
1073-6085
Publication state
Published
Issued date
2014
Peer-reviewed
Oui
Volume
56
Number
12
Pages
1100-1109
Language
english
Notes
Publication types: Journal Article Publication Status: ppublish
Abstract
Applications of genetic constructs with multiple promoters, which are fused with reporter genes and simultaneous monitoring of various events in cells, have gained special attention in recent years. Lentiviral vectors, with their distinctive characteristics, have been considered to monitor the developmental changes of cells in vitro. In this study, we constructed a novel lentiviral vector (FUM-M), containing two germ cell-specific promoters (Stra8 and c-kit), fused with ZsGreen and DsRed2 reporter genes, and evaluated its efficiency in different cells following treatments with retinoic acid and DMSO. Several cell lines (P19, GC-1 spg and HEK293T) were transduced with this vector, and functional capabilities of the promoters were verified by flow cytometry and quantitative RT-PCR. Our results indicate that FUM-M shows dynamic behavior in the presence and absence of extrinsic factors. A correlation was also observed between the function of promoters, present in the lentiviral construct and the endogenous level of the Stra8 and c-kit mRNAs in the cells. In conclusion, we recommend this strategy, which needs further optimization of the constructs, as a beneficial and practical way to screen chemical inducers involved in cellular differentiation toward germ-like cells.
Pubmed
Web of science
Create date
11/12/2014 18:03
Last modification date
20/08/2019 14:52
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