NR AXHV
AU Alden,J.; Grigoriadis,A.; Jat,P.; Collinge,J.; Klöhn,P.C.
TI Revertant Subclones Derived from Highly Susceptible Cells as a Tool for the Identification of Susceptibility Factors
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Oral Abstracts FC4.6
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Vortrag
AB Polymorphisms in the prion protein gene are well-know susceptibility factors of prion diseases. However, significant differences in incubation times for scrapie in mice with the same genotype indicate a role of prion protein-independent factors. To identify genetic factors that render cells susceptible to prions we subcloned the highly susceptible neuroblastoma clone N2aPK1 to isolate revertant clones. Three out of seven hundred clones (R2, R5 and R7) showed approximately a hundred-fold reduced rate of prion propagation as compared to the parental clone. This strategy for the isolation of resistant cells keeps epigenetic variations at a minimum as evidenced by the similarities of gene expression profiles between N2aPK1 subclones as compared to clones independently derived from neuroblastoma cells. To determine the impact of prion protein expression levels on susceptibility we transfected revertant clones with the mouse prion protein and pooled stably expressing clones after 10 days of antibiotic selection. Remarkably, enhanced levels of the prion protein (PrPc) as determined by PrPc surface expression levels were not associated with increased rates of prion propagation in revertant as well as in susceptible cells arguing that factors other than the prion protein are required for the cellular propagation of prions.
AD J. Alden, P. Jat, ,J. Collinge P.-C. Klohn, Institute of Neurology, MRC Prion Unit, UK; A. Grigoriadis, Ludwig Institute for Cancer Research, UK
SP englisch
PO Schottland