NR AXXG
AU Stobart,M.J.; Lamoureux,L.; Carpenter,M.; Knox,J.D.
TI RNAi-Mediated Knockdown of Genes Essential to Prion Protein Pathogenesis Enhances Neuroblastoma Resistance to PrP106-126 Cytotoxicity
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.159
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB The resistance of PrP knockout mice (prnp0/0) to prion infection has demonstrated that the protein product of the prnp gene, PrP, is essential to prion pathogenesis. Nonetheless, the molecular mechanisms resulting in prion-induced neurodegeneration remain poorly understood. We hypothesize that reduced expression of genes essential to prion pathogenesis will render neuronal cells resistant to PrP106-126 peptide mediated toxicity. Initial results targeting prnp mRNA demonstrate that the degree of prnp mRNA knockdown correlates to neuronal cell resistance to PrP106-126 exposure. Furthermore, the region of mRNA targeted and the structure of the inhibitor plays a role in determining the efficiency of mRNA knockdown. The knowledge gained from this pilot study was applied to the high throughput screening of a library of mRNA inhibitors. In this study, we have used complementary RNA inhibitors to reduce the levels of expression of specific mRNA. Multiple sites along the mRNA were targeted to ensure that at least one reduced mRNA expression sufficiently. Pure neuronal cell populations containing the targeted inhibitors were selected via antibiotic resistance and then were exposed to 80µM PrP106-126 peptide. Resistance to PrP106-126 induced cell death was determined by the ability of resistant cells to re-grow following challenge. PrP106-126 resistant inhibitor expressing clones permit identification of genes non-essential for host viability, but required for prion-induced cytotoxicity.
AD M. Stobart, The University of Manitoba, Medical Microbiology and Infectious Diseases, Canada; L. Lamoureux, J.D. Knox, Public Health Agency of Canada, Prion Disease Program, Canada; M. Carpenter, Public Health Agency of Canada, Division of Viral Diseases, Canada
SP englisch
PO Schottland