Tg(Prnp*M128*D177N)FFI-26Rchi

A cDNA derived from the a allele of the mouse prion protein gene (Prnpa) was modified to encode asparagine in place of aspartate at amino acid position 177 (D177N ) and methionine at position 128 (M128), orthologous respectively to the human D178N mutation and M129 polymorphism that together are associated with fatal familial insomnia (FFI). This cDNA was cloned into the expression vector MoPrP.Xho, which contains the mouse genomic Prnp sequence including exon 1, intron 1, and exon 2 joined directly to exon 3 in which the coding sequence has been replaced by an XhoI restriction endonuclease site, thus providing the promoter, 5'-untranslated region (5'-UTR), and 3'-UTR. Hemizygous mice of this transgenic line were determined by quantitative PCR to carry ~15 co-integrated copies of the transgene and by immunoblot analysis of brain protein extracts to express the transgene-encoded PRNP protein at approximately the same level as do wild-type mice from the endogenous gene. (J:235930)