A targeting construct was designed to insert a cre/ERT2 sequence, an SV40 polyA site, and a floxed neo selection cassette into the Aicda locus, replacing exon 2 and fusing with sequence encoding the first 4 amino acids, including 4 bases downstream of the exon 2 splice site. The splice acceptor site of Aicda intron 2-3 was mutated to prevent splicing events that could eliminate the cre/ERT2 sequence. E14.1 ES cells were electroporated with the linearized vector. Recombinant clones were transiently transfected with a cre-expressing vector to excise the neo cassette. (J:157741)