Smg6^tm1.1Tac

A loxP site, an FRT site flanked neomycin resistance gene cassette, cDNA sequence for exons 17-19, human GH gene poly(A) signal sequences, an F3 site flanked puromycin resistance gene cassette and a second loxP site were inserted into intron 16. Aspartic acid codons 1352 (GAT) and 1391 (GAT) in endogenous exons 18 and 19 were changed to alanine (GCC) (p.D1352A, p.D1391A). These mutations, in the PIN (PilT N terminus) nuclease domain of the encoded peptide, render the protein NMD-inactive. The neo and puro cassettes were removed through subsequent Flp-mediated recombination. This allele expresses wildtype peptides and only after Cre-mediated deletion of the exon 17-19 and pol(A) signal sequences will it express the mutated peptides. (J:342742)