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Caspase-11 Promoter-GFP Construct as a Dual Reporter of Cytotoxicity and Inflammation

机译:Caspase-11启动子-GFP构建为细胞毒性和炎症的双重报告者。

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摘要

Caspase-11 has been known as a dual regulator of apoptosis and inflammatory response. An unusual feature of caspase-11 is that its expression is induced by apoptotic or proinflammatory stimuli. Utilizing these unusual features of caspase-11, we have developed a simple and sensitive assay method to screen pro- or anti-apoptotic/inflammatory molecules. To develop this assay method, we generated a reporter construct where GFP expression is regulated by caspase-11 promoter. When several types of culturedcells were transfected with this reporter construct and subsequently treated with various apoptotic or proinflammatory molecules, expression of GFP by the activation of caspase-11 promoter was easily detected by fluorescence microscopy or spectro-fluorometry. In addition, a reduction of the GFP fluorescence was detected when an agent reported to suppress caspase-11 induction was applied. These results suggest that our reporter system can be used to screen pro- or anti-apoptotic/inflammatory molecules.
机译:Caspase-11被称为凋亡和炎症反应的双重调节因子。 caspase-11的一个不寻常特征是其表达是由凋亡或促炎性刺激诱导的。利用caspase-11的这些异常特征,我们开发了一种简单而灵敏的测定方法来筛选促凋亡或抗凋亡/炎症分子。为了开发这种测定方法,我们生成了一个报告子构建体,其中的GFP表达受caspase-11启动子调控。当几种类型的培养细胞用该报告基因构建体转染并随后用各种凋亡或促炎分子处理时,通过荧光显微镜或荧光分光光度法很容易检测到通过激活caspase-11启动子的GFP表达。另外,当应用据报道抑制caspase-11诱导的试剂时,检测到GFP荧光的减少。这些结果表明我们的报告系统可用于筛选促凋亡或抗凋亡/炎症分子。

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