Generation and analysis of conditional inducible transgene mice for the analysis of cFLIP in the skin

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The characterization of the function of genes in the organism has been revolutionized by the generation of transgenic animals that do not express the gene of interest or only express it in modified form. In the last decade the technology of inducible cell type-specific transgene or knockout animals has enabled the analysis of genes within different cell types. These Cre/loxP-based technologies will be used in the project to study the function of the caspase-8 regulator cFLIP in keratinocytes. cFLIP plays a malor role during transmission of signals activated by death receptors. cFLIP is crucial in the intracellular regulation of apoptosis, NF-kB-dependent inflammatory activation and, as recently discovered, in the regulation of necroptosis. cFLIP is not only able to block death receptor apoptosis signalling, but can also regulate the ripoptosome, a novel signalling platform recently described by our group. Conventional, but also Epidermis-specific cFLIP knockout animals are embryonically lethal, as we have shown in unpublished preliminary work. Thus it is the goal of the project to analyze animals in which the cFLIP locus can be inducibly deleted post partem. With these mice we will be able to study the function of cFLIP in murine keratinocytes in the context of different forms of cell death, or inflammatory activation of the epidermis. Our experimental goals will include in vivo as well as in vitro experiments. We will characterize death receptor-associated and intracellular complexes such as the ripoptosome under conditions of cFLIP loss. Thereby we will aim to find mechanistic insight into the role of cFLIP for the different processes of apoptosis, necroptosis, and inflammatory activation derived from epidermis-initiated signals.