AnNF-κBluciferasereporterconstructisstablyintegratedintothegenomeofCHO-K1cells.ThefireflyluciferasegeneiscontrolledbytheNF-κBresponseelementlocatedupstreamoftheTATApromoter.Followingactivationbystimulants,endogenousNF-κBtranscriptionfactorsbindtotheDNAresponseelementstoinducetranscriptionoftheluciferasegene.
TheNF-κB-luciferase/CHO-K1celllineissuitableformonitoringtheactivityofNF-κB
transcriptionfactorthroughluminescencereadout.).Thiscelllinerespondstohumancytokine
IL-1β,respondsmoderatelytohumanTNF,anddoesnotrespondtohumanIFN-&lamBDa;(2μg/ml).
Reducingtheamountofserumduringincubationperiodmayincreasethesensitivityto
cytokines.SinceCHO-K1cellsdonotexpressendogenoushumanproteins,thiscellline
providesanexcellentplatformtoenableexogenousexpressionofaproteinofinteresttostudy
itsdownstreameffectonNF-κBsignaling.
2.Railo,A.,et.al.(2008)Wnt-11signalingleadstodown-regulationoftheWnt/betacatenin,JNK/AP-1andNF-kappaBpathwaysandpromotesviabilityintheCHO-K1cells.ExpCellRes.314:2389-99
3.Murphy,S.H.,et.al.(2011)Tumorsuppressorprotein(p)53,isaregulatorofNF-κBrepressionbytheglucocorticoidreceptor.Proc.Natl.Acad.Sci.USA108:17117-17122
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