NR AUSX
AU Klose,R.; Wünsch,A.; Zakhartchenko,V.; Hao,R.; Yang,F.; Reichenbach,H.D.; Wenigerkind,H.; Mitko,K.; Weppert,M.; Wolf,E.
TI Efficient targeting of PRNP in bovine primary fibroblasts - basis for the generation of PrPc-deficient cattle for prion research
QU TSE-Forum, 6. Kongress - Nationale TSE-Forschungsplattform, Greifswald 26.6.-28.6.2006, Poster: Therapie THE-03
PT Konferenz-Poster
AB
Cattle lacking PrPc expression are required for the evaluation of the physiological functions of PrPc in bovine as well as for clarifying the role of PrPc in the pathogenesis of BSE. If PrPc-deficient cattle proof to be resistant to BSE as is expected from studies in Prnp-/- mice (Büeler et al., 1993), such cattle are very important for animal biotechnology, providing a source of biopharmaceutical products devoid the risk for BSE. Due to the lack of germ-line competent embryonic stem cells in livestock species, nuclear transfer using targeted donor cells is so far the only possibility to perform gene knock-outs in livestock species.
The functional inactivation of the bovine PRNP was performed in primary bovine fibroblasts by sequential gene targeting using two targeting vectors in which the entire coding sequence for PrPc was replaced by a promoter-less neomycin and hygromycin resistance gene, respectively. The second targeting was performed in primary PRNP+/- fibroblasts obtained from a PRNP+/- fetus after nuclear transfer.
Correct targeting in PRNP-/- fibroblasts was verified by Southern blot analysis. FACS analysis of fibroblasts obtained from a PRNP-/- fetus showed absence of PrPc. On PRNP+/- fibroblasts we detected reduced PrPc levels compared to wildtype. Nuclear transfer using PRNP-/- fibroblasts as nuclear donor cells and embryo transfer are continued to produce PrPc-deficient offspring.
AD Regina Klose, Annegret Wünsch, Valeri Zakhartchenko, Ru Hao, Feikun Yang, Horst-Dieter Reichenbach, Hendrik Wenigerkind, Katrin Mitko, Myriam Weppert, Eckhard Wolf, Institute of Molecular Animal Breeding and Biotechnology, University of Munich
SP englisch
PO Deutschland
EA Photo des in Greifswald gezeigten Posters, dessen starke Abweichung vom Tagungsband von der Autorin Klose mit seiner größeren Aktualität erklärt wurde
OR Tagungsband