Tln1^tm1.2Geno

A minigene consisting of the entire wild-type sequence from exons 41 to the terminal coding exon 59, followed by a polyadenylation sequence was used in the targeting vector. The minigene and a FRT-flanked neo-selection cassette were flanked with loxP sites and placed upstream of a mutated exon 42 sequence that substitutes the glutamate at amino acid 1770 to an alanine. Flp-mediated recombination removed the neomycin cassette. Germline cre-mediated recombination removed the minigene resulting in splicing on to the mutated exon 42 leading to transcription of the E1770A mutant. This mutation is predicted to abrogate binding with the FERM domain and to block autoinhibition. (J:271017)