NF-kBluciferasereporterconstructisstablyintegratedintothegenomeofHCT-116cells.The
fireflyluciferasegeneiscontrolledby4copiesofNF-κBresponseelementlocatedupstreamof
theTATApromoter.Followingactivationbystimulants,endogenousNF-κBtranscriptionfactors
bindtotheDNAresponseelementstoinducetranscriptionoftheluciferasegene.
TheNF-κB-luciferase/HCT-116celllineissuitableformonitoringtheactivityofNF-κBsignaling
inresponsetostimulantssuchasthecytokinesTNFandIL-1β,pathogen-associated
molecularpattern(PAMP)(i.e.flagellin)orendogenousdamage-associatedmolecularpattern
(DAMP)molecules(i.e.NOD1ligand)(seeapplicationreferences).Itisalsosuitablefor
establishingcell-basedscreensforinhibitorsthattargetspecificNF-κBstimulatingmolecules.
ThiscelllinecanbefurthermodifiedtoallowinvestigationofdownstreamNF-κBactivitiesasa
resultoftargetedgeneticmutation(s).
1.SamuelTet.al.(2014)VariableNF-Bpathwayresponsesincoloncancercellstreatedwithchemotherapeuticdrugs.BMCCancer14:599.
2.ArabiAet.al.(2012)ProteomicscreenrevealsFbw7asamodulatoroftheNF-Bpathway.NatureCommunication
3:976.3.ClemoNKet.al.(2008)BAG-1isup-regulatedincolorectaltumourprogressionandpromotescolorectaltumourcellsurvivalthroughincreasedNF-Bactivity.Carcinogenesis29:849.
4.TukhvatulinAIet.al.(2011)AnInVitroandInVivoStudyoftheabilityofNOD1LigandstoActivatetheTranscriptionalFactorNF-B.ActaNaturae3:77.
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