Gardner, DP; Byrne, GW; Ruddle, FH; Kappen, C; (1996) Spatial and temporal regulation of a lacZ reporter transgene in a binary transgenic mouse system. Transgenic Res , 5 (1) 37 - 48.
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The transgenic mouse system is a powerful tool for the study of gene function. However, when the analysis involves genes that are critical for the normal developmental process, the usefulness of transgenic mouse systems is limited (for review see Hanahan, 1989; Westphal and Gruss, 1989; Byrne et al., 1991). This is due to potential transgene interference with development in case of ectopic or high level expression. As a result, establishing permanent transgenic mouse lines expressing these types of genes has proven difficult. To circumvent these difficulties, a binary transgenic mouse system has been established, termed the Multiplex System (Byrne and Ruddle, 1989). This is a two-tiered gene activation system in which expression of the gene of interest occurs only in offspring carrying transgenes encoding both components: transactivator and transresponder. Transactivator lines contain the gene encoding the VP16 protein of herpes simplex virus. Transresponder lines harbour the gene of interest linked to the IE promoter which includes recognition sequences for the VP16 transactivator. Previously, the inducibility of a chloramphenicol acetyltransferase reporter gene in newborn offspring that carried both a transactivator and transresponder transgene (Byrne and Ruddle, 1989) has been shown. Moreover, it has been demonstrated that expression of the VP16 protein was not detrimental to development and that transactivation appeared to be tissue specific. Here, the potential of the system for the expression of transgenes in early mouse embryogenesis was examined, using the Escherichia coli beta-galactosidase gene as a reporter in the transresponder mouse strain. To direct expression of VP16, the murine Hoxc-8 promoter, which is known to be active during early development, was used. Embryos from crosses of transactivators to transresponders were isolated at different stages of development and stained for beta-galactosidase activity. Transactivation, as demonstrated by strong beta-galactosidase staining, could be detected as early as eight days of development. At all stages examined, the pattern of lacZ transresponder gene expression accurately reflected the activity of the Hoxc-8 promoter controlling VP16 expression. It is demonstrated that the Multiplex System can be used to express transresponder transgenes in a spatially and temporally defined manner in multiple cell types early during mouse embryogenesis.
|Title:||Spatial and temporal regulation of a lacZ reporter transgene in a binary transgenic mouse system.|
|Keywords:||Animals, Base Sequence, Embryonic and Fetal Development, Female, Gene Expression Regulation, Developmental, Herpes Simplex Virus Protein Vmw65, Homeodomain Proteins, Immediate-Early Proteins, Mice, Mice, Inbred C57BL, Mice, Inbred Strains, Mice, Transgenic, Models, Genetic, Molecular Sequence Data, Pregnancy, Promoter Regions, Genetic, Recombinant Proteins, Reproducibility of Results, Tissue Distribution, Transcriptional Activation, beta-Galactosidase|
|UCL classification:||UCL > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute of Cardiovascular Science|
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