KappaBle fluorescent reporter mice enable dynamic and low-background single-cell detection of NF-kB activity in vivo
Nuclear factor-{kappa}B (NF-{kappa}B) 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-{kappa}B 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-{kappa}B 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-{kappa}B, the latter have the potential to allow the analysis of NF-{kappa}B 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 knockin NF-{kappa}B reporter (KappaBle) mice containing a destabilized EGFP, which showed sensitive, dynamic, and faithful monitoring of NF-{kappa}B activity at the single-cell level of various cell types during inflammatory and infectious diseases.