Department of Biology, Institute of Molecular Health Sciences, ETH, Zurich, Switzerland.
Department of Biology, Institute of Microbiology, ETH, Zurich, Switzerland.
Mucosal Immunol. 2022 Apr;15(4):656-667. doi: 10.1038/s41385-022-00525-8. Epub 2022 May 19.
Nuclear factor-κB (NF-κB) is a transcription factor with a key role in a great variety of cellular processes from embryonic development to immunity, the outcome of which depends on the fine-tuning of NF-κB activity. The development of sensitive and faithful reporter systems to accurately monitor the activation status of this transcription factor is therefore desirable. To address this need, over the years a number of different approaches have been used to generate NF-κB reporter mice, which can be broadly subdivided into bioluminescence- and fluorescence-based systems. While the former enables whole-body visualization of the activation status of NF-κB, the latter have the potential to allow the analysis of NF-κB activity at single-cell level. However, fluorescence-based reporters frequently show poor sensitivity and excessive background or are incompatible with high-throughput flow cytometric analysis. In this work we describe the generation and analysis of ROSA26 knock-in NF-κB reporter (KappaBle) mice containing a destabilized EGFP, which showed sensitive, dynamic, and faithful monitoring of NF-κB transcriptional activity at the single-cell level of various cell types during inflammatory and infectious diseases.
核因子-κB（NF-κB）是一种转录因子，在从胚胎发育到免疫等多种细胞过程中发挥关键作用，其结果取决于 NF-κB 活性的精细调节。因此，需要开发灵敏且准确的报告系统来精确监测该转录因子的激活状态。为了满足这一需求，多年来已经使用了许多不同的方法来生成 NF-κB 报告小鼠，这些方法大致可分为基于生物发光和荧光的系统。虽然前者能够实现 NF-κB 激活状态的全身可视化，但后者有可能允许在单细胞水平上分析 NF-κB 活性。然而，基于荧光的报告器通常显示出较差的灵敏度和过度的背景，或者与高通量流式细胞术分析不兼容。在这项工作中，我们描述了 ROSA26 基因敲入 NF-κB 报告（KappaBle）小鼠的生成和分析，该小鼠含有不稳定的 EGFP，可在炎症和感染性疾病期间各种细胞类型的单细胞水平上灵敏、动态和准确地监测 NF-κB 转录活性。
