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

用于分析皮肤中 cFLIP 长亚型和短亚型的条件和诱导转基因小鼠模型的生成和分析

• 批准号: 397983964
• 负责人: Dr. Diana Panayotova Dimitrova, Ph.D.
• 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: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/397983964?language=en
• 关键词: Generation; analysis; conditional; inducible; transgenic

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.

在过去的几十年里，用于研究基因功能的小鼠模型的生成技术取得了显着进步。cre/lox系统用于产生组织特异性和诱导性基因缺失，是在空间和时间上控制基因活性的有力工具。在本项目中，我们旨在分析cFLIP长（cFLIPL）和cFLIP短（cFLIPS）异构体在体内皮肤中的作用。这将通过生成和分析具有选择性表达cFLIPL或表皮细菌人工染色体（BAC）表达的cFLIPL亚型的小鼠模型来实现。我们的模型允许他莫昔芬诱导（CreERTam） cre介导的内源性cFLIP位点缺失。我们最近确定了cFLIP的两种亚型在调节细胞内细胞死亡决策过程中的关键和差异作用，这些过程控制着细胞内中心信号平台-核核糖体中的细胞凋亡或坏死坏死。我们还表明，体内表皮特异性的完整cFLIP缺失会导致暴发性炎症和tnf依赖性细胞凋亡。然而，个体cFLIP异构体在体内皮肤中的作用仍然完全未知。利用cfllip -/-和cflps -/-转基因动物，我们将能够详细研究单个cflp亚型对胞内平台（如Ripoptosome）及其下游信号凋亡和necroptosis对体内自发和诱导细胞死亡的控制的影响。我们将研究单个cFLIP亚型缺失后炎症性皮肤病的动力学、程度和病程。我们的实验设置将包括体内和体外实验。因此，我们的目标是确定通过调节细胞凋亡、坏死和炎症等重要过程来维持皮肤稳态的cFLIP亚型的控制机制。