A loxP site was inserted upstream of exon 1 and an FRT site flanked neomycin resistance gene cassette and a second loxP site downstream of exon 2. Also inserted downstream of exon 2 was a cDNA for the endogenous gene where a point mutation was engineered to change aspartate codon 34 into an asparagine codon (p.D34N or p.Asp34Asn). The neo cassette was removed through subsequent flp-mediated recombination. This leaves a conditional-ready allele with the entire gene (exons 1-2) floxed. Only after cre-mediated deletion of the endogenous gene will the mutated cDNA, which codes for a kinase-dead version of the protein, be expressed. (J:259970)
Legend:
cx: complex: > 1 genome feature ot: other: hemizygous, indeterminate,... (F): Female
(M): Male
N: normal phenotype
(#): related diseases count