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Chimeric prion protein expression in cultured cells and transgenic mice
Author(s) -
Scott Michael R.,
Köhler Ruth,
Foster Dallas,
Prusiner Stanley B.
Publication year - 1992
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560010804
Subject(s) - biology , transgene , microbiology and biotechnology , fusion protein , mutagenesis , gene , monoclonal antibody , chimeric gene , genetically modified mouse , transformation (genetics) , scrapie , expression vector , transfection , epitope , gene expression , virology , antibody , mutation , genetics , prion protein , recombinant dna , medicine , disease , pathology
The efficient expression of exogenous prion protein (PrP) molecules in mouse neuroblastoma cells that are chronically infected with murine scrapie prions (ScN 2 a cells; Butler, D.A., et al., 1988, J. Virol. 62 , 1558–1564) and in transgenic mice is described. This technology allows investigation of the PrP molecule for structural regions involved in determining species specificity, as well as ablation experiments designed to address the functionality of particular regions of the PrP molecule. Previous reports demonstrated that the PrP gene specifies the host range for susceptibility of transgenic animals to prions (Scott, M., et al., 1989, Cell 59 , 847–857; Prusiner, S.B., et al., 1990, Cell 63 , 673–686). Consistent with these results, we showed that Syrian hamster (SHa) PrP is ineligible for efficient conversion to PrP Sc in ScN 2 a cells. By constructing a series of chimeric mouse (Mo)/SHaPrP genes, we developed an epitopically tagged functional variant of the MoPrP gene, which can efficiently form protease‐resistant PrP molecules upon expression in ScN 2 a cells. The presence of a defined epitope for an SHa‐specific monoclonal antibody allows the products of this chimeric gene to be discriminated from endogenous MoPrP and creates a useful reagent for exploring structure/function relationships via targeted mutagenesis. In addition, we developed a transgenic mouse expression vector by manipulation of an SHaPrP cosmid clone. This vector permits the efficient expression of foreign PrP genes in the brains of transgenic animals, enabling pathological consequences of in vitro mutagenesis to be studied.