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Construction of RNAi Expressing Vector Targeting the Goat Prion Protein Gene (PRNP)

Author(s): Yongjun Li | Wenting Li | Taikun Li | Guangjing Zhang | Jiahui Zhang | Dejun Ji

Journal: Journal of Animal and Veterinary Advances
ISSN 1680-5593

Volume: 11;
Issue: 8;
Start page: 1208;
Date: 2012;
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Keywords: RNA interference | siRNA | prion protein gene | goat | antagonistic | protein

To explore the effect of RNA interference on the suppression of goat prion protein gene (PRNP), small interfering RNA (siRNA) expressing vectors pSilencer4.1-siRNA1 and a negative control pSilencer4.1-siRNA2 and the eukaryotic expressing vector pSG5-prion expressing parts of goat prion protein gene were designed and constructed. The recombinants were introduced into Human Embryonic Kidney (HEK) 293T cells mediated by GeneTran. RT-PCR was carried out to detect the expression of prion protein gene. The results showed that RNAi effects induced by pSilencer4.1-siRNA1 could successfully deplete the mRNA expression of goat prion protein gene in 293T cells 48 h after transfection with both vectors compared to the negative control. The findings demonstrated that pSilencer4.1-siRNA1 was constructed successfully to transfer antagonistic substance against goat prion protein gene.
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