PUBLICATION

Application of RNAi Technology to the Inhibition of Zebrafish GtHalpha, FSHbeta, and LHbeta Expression and to Functional Analyses

Authors
Huang, W.T., Hsieh, J.C., Chiou, M.J., Chen, J.Y., Wu, J.L., and Kuo, C.M.
ID
ZDB-PUB-080722-6
Date
2008
Source
Zoological science   25(6): 614-621 (Journal)
Registered Authors
Chen, Jyh-Yih, Huang, Wei-Tung, Wu, Jen-Leih
Keywords
zebrafish, RNA interference, knockdown, gonadotropin alpha, GtHα, common α, follicle-stimulating hormone beta, FSHβ, luteinizing hormone beta, LHβ
MeSH Terms
  • Reverse Transcriptase Polymerase Chain Reaction
  • RNA Interference*
  • Gonadotropins, Pituitary/metabolism*
  • Gonadotropins, Pituitary/pharmacology
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
  • Microinjections
  • Genetic Vectors/genetics
  • Animals
  • Green Fluorescent Proteins/metabolism
  • RNA/metabolism
  • DNA Primers/genetics
  • Blotting, Southern
  • Embryonic Development/drug effects*
  • Embryonic Development/genetics
(all 15)
PubMed
18624572 Full text @ Zool. Sci.
Abstract
Zebrafish (Danio rerio) were used as a model fish, and the technique of RNA interference (RNAi) was employed to knockdown three subunits of the gonadotropin alpha (GtHalpha, common alpha), follicle-stimulating hormone beta (FSHbeta), and luteinizing hormone beta (LHbeta) genes. Three short-hairpin RNA (shRNA) expression vectors and three mismatched shRNA expression vectors as controls for each subunit gene were constructed, and the depression efficiency was tested in vivo by microinjection; the RNA or protein expression levels of the GtH genes were monitored by RT-PCR, Southern blotting, and green fluorescent protein (GFP) analyses. Expression of GtH mRNA was obviously and more efficiently depressed by GtHalpha RNAi expression compared with the other two subunits. A GtHalpha morpholino analysis showed that the GtHalpha morpholino led to suppression of embryonic development and the production of embryonic mutants as a result of an injection of GtHalpha -shRNA. Taken together, these results show that GtHalpha-shRNA, which more efficiently targets RNAi, may have an essential role in the further development of sterility technology of transgenic fish for biosafety purposes.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
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Citation
Huang, W.T., Hsieh, J.C., Chiou, M.J., Chen, J.Y., Wu, J.L., and Kuo, C.M. (2008) Application of RNAi Technology to the Inhibition of Zebrafish GtHalpha, FSHbeta, and LHbeta Expression and to Functional Analyses. Zoological science. 25(6):614-621.