NR AWPQ
AU Vana,K.; Ludewigs,H.; Ruhmann,M.; Rank,C.; Messow,D.; Mitteregger,G.; Nikles,D.; Weiss,S.
TI LRP-downregulation transfert using siRNA delivery via lentiviral vector gene
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 THE-20
PT Konferenz-Poster
AB Transmissible spongiform encephalopathies (TSEs) are neurodegenerative diseases, which include Scrapie in sheep, BSE in cattle and CJD in humans. Prions, the causative agents of TSEs, are known to interact with the cellular prion protein (PrPc) by inducing conformational changes. The 37 kDa/ 67 kDa laminin receptor (LRP/ LR) acts as the cell surface receptor for the cellular PrP (1) and the infectious prion protein (2,3). It has been shown that LRP/ LR is essential for PrPsc propagation in neuronal cells (4). The accumulation of PrPsc in scrapie-infected neuronal cells (N2aSc+) has been prevented by transfection with small interfering (si) RNAs specific for the LRP mRNA (4). These results demonstrate the necessity of the laminin receptor for the PrPsc propagation in cultured cells. Vector-based application of siRNAs circumvents the transient effect of downregulation of gene expression and allows persistent suppression and therefore analysis of loss-of-function-phenotypes that develop over longer periods of time. Transduction of recombinant HIV-based lentiviral vectors expressing siRNAs directed against defined regions of the LRP mRNA resulted in reduction of both PrPres and LRP levels in scrapie-infected neuronal cells. To further enlighten the role of LRP/LR in prion diseases, injection of recombinant LRP-specific RNA interference (RNAi) lentiviral particles into mice using an intracerebral (i.c.) or intraperitoneal (i.p.) route was performed. Western blot analysis of the cortical brain area of mice intracerebrally injected with lentiviral vectors expressing siRNAs directed against the LRP mRNA showed a downregulation of the 67kDa LR. Subsequent prion inoculation in these mice will prove whether the knockdown of LRP/LR by RNAi might prolong the onset of TSE or even prevent prion disease. (1) Gauczynski et al. (2001) EMBO J. 20, 5863-5875. (2) Morel et al. (2005) Am. J. Path., 167, 10331042 (2005). (3) Gauczynski et al., (2006) J. infect. Dis. in press. (4) Leucht et al. (2003) EMBO rep 4, 290-295.
AD K. Vana, H. Ludewigs, M. Ruhmann, C. Rank, D. Messow, D. Nikles, S. Weiss: Genzentrum - Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 Munich, Germany; G. Mitteregger: Zentrum für Neuropathologie und Prionforschung der LMU München, Feodor-Lynen-Str. 23, 81377 München, Germany; E-mail: weiss@lmb.uni-muenchen.de
SP englisch
PO Italien