Generation and analysis of conditional and inducible transgenic mouse models for analysis of cFLIP long and short isoforms in skin

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In the last decades, technologies for the generation of mouse models for studying gene function, have advanced significantly. The cre/lox system used for the generation of tissue specific and inducible gene deletion represents a powerful tool for the control of gene activity in space and time. In the proposed project we aim to analyze the role of cFLIP long (cFLIPL) and cFLIP short (cFLIPS) isoforms in skin in vivo. This will be achieved by generation and analyses of mouse models with selective expression of either cFLIPL or cFLIPS isoforms expressed from a bacterial artificial chromosome (BAC) in the epidermis. Our model allows the Tamoxifen-induced (CreERTam) Cre-mediated deletion of the endogenous cFLIP locus. We have recently identified the critical and differential role of both isoforms of cFLIP in the regulation of intracellular cell death decision processes that govern either apoptosis or necroptosis within the central intracellular signaling platform - Ripoptosome. We have also shown that epidermal-specific loss of complete cFLIP in vivo leads to a fulminant inflammatory and TNF-dependent apoptosis. However the role of the individual cFLIP isoforms in the skin in vivo remains completely unidentified. With cFLIPL-/- und cFLIPS-/- transgenic animals we will be able to study in details the impact of the individual cFLIP isoforms on the control of intracellular platforms such as the Ripoptosome and their downstream signals apoptosis and necroptosis for spontaneous and induced cell death in vivo. We will study the kinetics, extent and course of inflammatory skin disease upon deletion of individual cFLIP isoforms. Our experimental settings will include in vivo as well as in vitro experiments. Thereby we will aim to define control mechanisms of cFLIP isoforms for the maintenance of skin homeostasis through regulation of important processes such as apoptosis, necroptosis and inflammation.