A targeting vector was designed to place a loxP-flanked thymidine kinase/neomycin phosphotransferase expression (tk/neo) cassette upstream of exon 3, and a third loxP site just downstream of exon 5 of the targeted gene. Correctly targeted ES cells were transiently transfected with a cre expressing plasmid to remove the selection cassette (generating the Gata4-loxP allele; with one loxP site upstream of exon 3 and one loxP site downstream of exon 5). (details provided by SA Duncan, personal communication.) (J:92434)