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Insertion vector IV-3KO was used to insert a cassette containing an FRT-flanked neo with a 3' loxP site and a copy of exon 4 with a second loxP site downstream. The duplication of exon 4 was predicted to shift the reading frame and prevent formation of the protein. Crossing with mice expressing FLP recombinase removed the neomycin resistance gene. RT-PCR failed to detect transcript containing the wild-type allele or the duplicate exon. Northern blot, Western blot and immunohistochemical analyses confirmed absence of transcript and protein in mutants, respectively. (J:103831)