NR AWOL
AU Stobart,M.J.; Carpenter,M.; Knox,J.D.
TI Targeted knock-down of mRNA essential to prion pathogenesis reduces sensitivity to PrP106-126 cytotoxicity
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PA-55
PT Konferenz-Poster
AB Due to the interspecies conveyance and the terminal disease caused by the infectious prion protein, there has been much interest in identifying key molecular players in disease progression. To date, the only known gene product essential to prion pathogenesis is the PrP protein, the product of the prnp gene. Previous studies involving PrP knockout mice (prnp0/0) have shown that challenge with infectious PrP-containing homogenate fails to lead to disease, and that neurons derived from these knockout mice are resistant to the cytotoxicity of the PrP106-126 peptide in vitro. These observations parallel experiments that demonstrate that over expression of the PrP protein in animals succumb to disease at a faster rate than those expressing normal levels of PrP. The knockdown of a factor nonessential for viability of the host, but required for existence of the pathogen or its capacity to initiate pathogenesis, will make cells less susceptible to pathogen mediated toxicity. In this study, we have used inhibitors specifically targeting prnp mRNA for knockdown to observe the protective effect significantly reduced levels of prnp mRNA gene expression confers upon neuronal cells exposed to the cytotoxic PrP106-126 peptide. Cell populations were generated with the targeted inhibitors, which either lacked or possessed a modification that permitted enhanced prnp mRNA interaction as well as knockdown function. To determine whether the location targeted along the prnp mRNA plays a role in knockdown efficiency, four sites were chosen. These targeted inhibitor-containing cell populations were exposed to 80 M PrP106-126 peptide and their resistance to PrP106-126 induced cell death was determined by flow cytometry analysis. The results demonstrate that significant knockdown of prnp mRNA reduces neuronal cell sensitivity to PrP106-126 exposure. Our results also demonstrate that the site along the mRNA targeted for knockdown does play a role in determining the efficiency of mRNA knockdown and therefore the extent of PrP106-126 resistance.
AD M.J. Stobart, J.D. Knox: Faculty of Medicine, Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Canada, and Public Health Agency of Canada, Division of Host Genetics and Prion Diseases, Canadian Science Center for Human and Animal Health, Winnipeg, Canada; M. Carpenter: Faculty of Medicine, Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, Canada, and Public Health Agency of Canada, Division of Viral Diseases, Canadian Science Center for Human and Animal Health, Winnipeg, Canada
SP englisch
PO Italien