Authors: Shaohua Wang Xiaoqing Lv Kun Zhang Tonghui Lin Xiaofang Liu Jing Yuan Yunping Dai Ning Li
Publish Date: 2009/10/11
Volume: 37, Issue: 7, Pages: 3193-3198
Abstract
PRNP is the gene encoding prion protein whose misfolded and βsheetrich isoform is the infectious agent of transmissible spongiform encephalopathy TSE TSE also called prion diseases cause fatal neurodegenerative and transmissible disorders in human and animals Among these diseases bovine spongiform encephalopathy BSE has tremendous impact on economy and human health in the world In the present study we hypothesize suppression of the PRNP gene expression could raise resistance to BSE in cattle by using vectorbased small interfering RNA siRNA expression systems Therefore the objective was to screen effective DNAencoding short hairpin RNAs shRNAs which could knockdown the PRNP gene expression in bovine fibroblast cells Human U6 promoter was employed to drive shRNA transcription from the DNA vector and seven shRNAs that designed to target coding region and 3′ untranslated region of the PRNP gene were selected Four out of seven shRNAs tested were found to be effective in inhibiting the PRNP gene expression and the most significant suppression level was as much as 629 evidenced by realtime RTPCR Furthermore the protein abundance was obviously reduced compared to the control Overall the present study demonstrated that vectorbased siRNA expression systems is an efficient approach to knockdown the PRNP gene expression in bovine fibroblast cells and thereby provide donor cells for somatic cell nuclear cloning to produce cattle that is resistant to prion related diseases
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