Paper

KappaBle fluorescent reporter mice enable dynamic and low-background single-cell detection of NF-kB activity in vivo

Kopf, Manfred, Ruelicke, Thomas, Ampenberger, Franziska, Tortola, Luigi, Rosenwald, Esther, Kisielow, Jan, Heer, Sebastian

2021

Overview

Nuclear factor-kB (NF-kB) 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-kB 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-kB 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-kB, the latter have the potential to allow the analysis of NF-kB 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-kB reporter (KappaBle) mice containing a destabilized EGFP, which showed sensitive, dynamic, and faithful monitoring of NF-kB activity at the single-cell level of various cell types during inflammatory and infectious diseases. Competing Interest Statement The authors have declared no competing interest.

• Publisher: Cold Spring Harbor Laboratory Press
• Subject: Bioluminescence / Embryogenesis / Flow cytometry / Infectious diseases / NF-κB protein / Transcription activation / Transcription factors