Selection for mutations in the cDNAs of transgenic mice upon expression of an embryonic lethal protein

The generation of transgenic mouse models to study in vivo functions of specific proteins has become common practice. In addition, PCR technology allows efficient and rapid identification of founder mice by the analysis of tail tip DNA. Whilst the DNA construct used in the microinjection of one-cell-stage embryos is usually sequenced it is not common practice to sequence the PCR product once the transgene has been inserted into the mouse genome. In this report we describe why sequencing of inserted transgenes is important. Upon generation of transgenic mice expressing a splice variant of MDM2, MDM2-A, three of four founders contained mutations within the Mdm2-a cDNA sequence. The observation that selection against expression of wild-type MDM2-A resulted in the generation of mice expressing mutant transgenes highlights the importance of sequencing the transgenes of founder mice.