title: GFP-p65 knock-in mice as a tool to study NF-kappaB dynamics in vivo.
abstract: The nuclear factor kappaB (NF-kappaB) signaling pathway regulates immune and inflammatory responses and is implicated in the pathogenesis of multiple diseases. The principal mechanism controlling NF-kappaB activation depends on the association of NF-kappaB transcription factor dimers with IkappaB molecules, which prevents the accumulation of NF-kappaB in the nucleus and the activation of target gene transcription. Monitoring the nucleocytoplalsmic shuttling of NF-kappaB factors is a reliable method to study the dynamic regulation of NF-kappaB activity. Here, we generated knock-in mice expressing a fusion protein between the green fluorescent protein (GFP) and the p65/RelA NF-kappaB subunit (GFP-p65) from the endogenous p65 genomic locus. Homozygous GFP-p65 mice developed normally and showed normal NF-kappaB activation, demonstrating that the GFP-p65 fusion protein functionally substitutes for wild-type p65. Live imaging of primary cells from GFP-p65 mice allowed real-time monitoring of p65 nucleocytoplasmic shuttling upon NF-kappaB activation. Therefore, the GFP-p65 knock-in mice constitute an invaluable tool for studying the dynamic regulation of NF-kappaB.
authors.affiliation: Institute for Genetics, Centre for Molecular Medicine, and Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Germany.
authors.forename: Rossana
authors.initials: R
authors.lastname: De Lorenzi
authors.forename: Ralph
authors.initials: R
authors.lastname: Gareus
authors.forename: Sven
authors.initials: S
authors.lastname: Fengler
authors.forename: Manolis
authors.initials: M
authors.lastname: Pasparakis
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D055879:Gene Knock-In Techniques / Q000379:methods*; D018390:Gene Targeting / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D015151:Immunoblotting; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D051996:Transcription Factor RelA; D049452:Green Fluorescent Proteins