NR AOWW
AU Milhavet,O.; Casanova,D.; Lehmann,S.
TI Use of Short Interfering RNA to Establish the Function of the Prion Protein
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-65
PT Konferenz-Poster
AB RNA interference allows sequence-specific, post-transcriptional gene silencing initiated by double-stranded RNA that is homologous in sequence to the silenced gene. Recently, 21-nucleotide small interfering RNA dimers (siRNA) have been shown to specifically suppress the expression of endogenous genes in mammalian cells. The development of this method provides a powerful tool for the study of gene function in mammalian cells. We have used siRNAs targeting the prion protein (PrP) in cell lines, primary neurons in culture and in neural stem cells to study the physiological function of PrP and its alterations during infection. Several 21-nucleotide siRNAs dimers were designed to knock-down PrP and their efficiency was confirmed by western blot and/or immunofluorescence analyses. Levels of PrP were successfully reduced in primary hippocampal neurons and in a neural precursor cell line. Silencing of PrP using siRNA in mouse neural stem cell cultures was also established. Specific knockdown of PrP using siRNAs should allow us to establish functions of PrP in physiological and pathological processes. This technique, an easier and faster approach than gene knockouts, is proving to be a powerful tool for establishing the functions of specific proteins in neural cells.
AD Ollivier Milhavet, Danielle Casanova, Sylvain Lehmann, IGH-CNRS UPR1142, Montpellier, France
SP englisch
PO Deutschland