PUBLICATION
The Transgenic Zebrafish Display Fluorescence Reflecting the Expressional Dynamics of Dihydrofolate Reductase
- Authors
- Chang, W.N., Chi, W.Y., Kao, T.T., Tsai, J.N., Liu, W., Liang, S.S., Chiu, C.C., Chen, B.H., Fu, T.F.
- ID
- ZDB-PUB-170330-12
- Date
- 2017
- Source
- Zebrafish 14(3): 223-235 (Journal)
- Registered Authors
- Chang, Wen-Ni, Fu, Tzu-Fun, Kao, Tseng-Ting
- Keywords
- dihydrofolate reductase, fluorescence transgenic embryos, folate, one-carbon metabolism, promoter, transcriptional regulation, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/growth & development
- Animals, Genetically Modified/metabolism*
- Embryo, Nonmammalian/metabolism*
- Fluorescence
- Gene Expression Regulation, Enzymologic*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism*
- HeLa Cells
- Humans
- Promoter Regions, Genetic
- Sp1 Transcription Factor/genetics
- Sp1 Transcription Factor/metabolism
- Tetrahydrofolate Dehydrogenase/genetics*
- Tetrahydrofolate Dehydrogenase/metabolism
- Zebrafish/genetics
- Zebrafish/growth & development
- Zebrafish/metabolism*
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 28350247 Full text @ Zebrafish
Citation
Chang, W.N., Chi, W.Y., Kao, T.T., Tsai, J.N., Liu, W., Liang, S.S., Chiu, C.C., Chen, B.H., Fu, T.F. (2017) The Transgenic Zebrafish Display Fluorescence Reflecting the Expressional Dynamics of Dihydrofolate Reductase. Zebrafish. 14(3):223-235.
Abstract
Dihydrofolate reductase (DHFR) reduces folic acid and recycles dihydrofolate generated during dTMP biosynthesis to tetrahydrofolate. DHFR is upregulated in rapidly proliferating cells and hence a favored target of antifolate drug against cancers, autoimmune diseases, and microbial infections. However, increased expression of dhfr contributed to the often emerging drug resistance and impeded the therapeutic efficacy of antifolate drugs. Therefore, comprehensive knowledge on the expressional control of dhfr becomes crucial. We generated two zebrafish transgenic lines, Tg(zdhfr+91:EGFP) and Tg(zdhfr+79:EGFP), which express green fluorescent protein driven by two zebrafish dhfr promoter fragments separately. The fluorescence intensity displayed in these transgenic embryos recapitulated the expressional dynamics of endogenous dhfr and reflected changes in dhfr mRNA and protein levels. The fluorescence intensity of these transgenic embryos was responsive to both genetic and environmental factors potentially modulating dhfr promoter activity. Sequence analyses revealed partial conservation on the landscape of transcription factor arrangement between zebrafish and human dhfr promoters. A noncanonical and inhibitory Sp1 site was identified 170 base-pair upstream to the conserved Sp1 site in close proximity to the translation initiation codon. Our results supported the potential use of these transgenic embryos for studying the expressional dynamics of dhfr and preliminary screening for dhfr promoter modulators.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping