Gene-trap-based target site for Cre-mediated transgenic insertion

There is an increasing need for tissue-specific gene expression regulatory elements to study normal and disease development in the mouse. However, the cloning and characterization of these elements are time-consuming and costly. Thus, there is a particular need to be able to identify gene expression patterns without having to clone the promoter elements. Gene-trap strategies identify expression patterns assigned for endogenous genes using reporters, such as LacZ (Gossler et al., 1989; Skarnes, 1990) or green fluorescent protein (GFP) (Ishida and Leder, 1999; Zheng and Hughes, 1999). The gene-trap vector randomly inserts into the genome and "steals" regulatory elements for the reporter. Here we describe an improved gene-trap strategy, which allows an efficient Cre recombinase-mediated insertion of any transgene into the trapped loci as a post-integrational modification and links the expression of the transgene to that of the reporter.