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

由于转基因动物的产生，对生物体中基因功能的描述已经发生了革命性的变化，这些转基因动物不表达感兴趣的基因或只以修饰的形式表达它。在过去的十年中，诱导细胞类型特异性转基因或敲除动物技术使不同细胞类型内的基因分析成为可能。这些基于Cre/ loxp的技术将在该项目中用于研究caspase-8调节因子cFLIP在角质形成细胞中的功能。cFLIP在死亡受体激活的信号传递过程中起着重要作用。cFLIP在细胞内调节细胞凋亡、nf - kb依赖性炎症激活以及最近发现的坏死性坏死中起着至关重要的作用。cFLIP不仅可以阻断死亡受体的凋亡信号，还可以调控核糖体，这是我们小组最近描述的一种新的信号传导平台。正如我们在未发表的初步工作中所显示的那样，常规的，但也有表皮特异性的cFLIP敲除动物是胚胎致死的。因此，该项目的目标是分析在动物中可以诱导地删除cFLIP位点的动物。通过这些小鼠，我们将能够在不同形式的细胞死亡或表皮炎症激活的背景下研究cFLIP在小鼠角质形成细胞中的功能。我们的实验目标将包括体内和体外实验。我们将描述死亡受体相关和细胞内复合物，如在cFLIP丢失的情况下的核溶酶体。因此，我们将致力于发现cFLIP在表皮启动信号引发的细胞凋亡、坏死下垂和炎症激活等不同过程中的作用机制。

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 Conventional, but also Epidermis-specific cFLIP knockout animals are embryonically lethal, as we have shown in unpublished preliminary work.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.