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

用于分析皮肤中 cFLIP 的条件诱导转基因小鼠的生成和分析

• 批准号: 229783597
• 负责人: Dr. Diana Panayotova Dimitrova, Ph.D., since 4/2016
• 金额: --
• 依托单位: Klinik für Dermatologie und Allergologie - Hautklinik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/229783597?language=en
• 关键词: Generation; analysis; conditional; inducible; transgene

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 Conventional, but also Epidermis-specific cFLIP knockout animals are embryonically lethal, as we have shown in unpublished preliminary work.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

通过产生不表达感兴趣的基因或仅以修饰形式表达它的转基因动物，生物体中基因功能的表征已经发生了革命性的变化。在过去的十年中，诱导型细胞类型特异性转基因或敲除动物的技术已经使得能够分析不同细胞类型内的基因。这些基于Cre/loxP的技术将用于研究角质形成细胞中caspase-8调节剂cFLIP的功能。cFLIP在死亡受体激活的信号传递过程中起重要作用。cFLIP在细胞凋亡、NF-κ B依赖性炎症激活的细胞内调节中以及最近发现的在坏死性凋亡的调节中是至关重要的。cFLIP不仅能够阻断死亡受体凋亡信号，而且还可以调节ripoptosome，这是我们小组最近描述的一种新的信号平台。常规的，但也表皮特异性cFLIP敲除动物是胚胎致死的，正如我们在未发表的初步工作中所示。因此，该项目的目标是分析其中cFLIP基因座可以在模式后诱导缺失的动物。通过这些小鼠，我们将能够研究cFLIP在不同形式的细胞死亡或表皮炎症活化的背景下在鼠角质形成细胞中的功能。我们的实验目标将包括在体内以及在体外实验。我们将表征死亡受体相关的和细胞内的复合物，如在cFLIP丢失的条件下的ripoptosome。因此，我们的目标是找到cFLIP的作用机制的洞察不同的过程中的细胞凋亡，坏死性凋亡，和炎症激活来自表皮启动的信号。