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25 June 2008 Application of RNAi Technology to the Inhibition of Zebrafish GtHα, FSHβ, and LHβ Expression and to Functional Analyses
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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 (GtHα, common α), follicle-stimulating hormone beta (FSHβ), and luteinizing hormone beta (LHβ) 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 GtHα RNAi expression compared with the other two subunits. A GtHα morpholino analysis showed that the GtHα morpholino led to suppression of embryonic development and the production of embryonic mutants as a result of an injection of GtHα -shRNA. Taken together, these results show that GtHα-shRNA, which more efficiently targets RNAi, may have an essential role in the further development of sterility technology of transgenic fish for biosafety purposes.

Wei-Tung Huang, Jung-Chen Hsieh, Ming-Jyun Chiou, Jyh-Yih Chen, Jen-Leih Wu, and Ching-Ming Kuo "Application of RNAi Technology to the Inhibition of Zebrafish GtHα, FSHβ, and LHβ Expression and to Functional Analyses," Zoological Science 25(6), 614-621, (25 June 2008). https://doi.org/10.2108/zsj.25.614
Received: 5 November 2007; Accepted: 1 March 2008; Published: 25 June 2008
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