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Signosis/GR Luciferase Reporter MDA-MB-453 Stable Cell Line/SL-0009-NP/1 Ea
Description:
GRResponsiveLuciferaseReporterMDA-MB-453StableCellLineisderivedfromhumanbreastcancer,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheGRresponseelement. Thiscelllineisanidealcellularmodelformonitoringtheactivation ofGlucocorticoidReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
TheGlucocorticoidreceptor(GR)isamajorplayerindevelopment,metabolismandimmuneresponse. Whenactivatedbystimuli,GRtranslocatesfromthecytoplasmintothenucleus,andthenbindsaDNArecognitionsitetoregulategeneexpression. Inaddition,activatedGRcantransrepressothertranscriptionfactorsthathavebeenmisregulatedincancerandotherdiseases.
ThecelllinewasestablishedbytransfectionofaGRfireflyluciferasereportervectoralongwithG418expressionvectorfollowedbyG418selection.TheG418resistantclonesweresubsequentlyscreenedforDEX-inducedluciferaseactivity.Theclonewiththehighestfoldinduction(38fold)wasselectedandexpandedtoproducethisstablecellline.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisofGRpathwayreporterMDA-MB-453celllineinresponsetostimuli. Thecellswereseededona96-wellplateforovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout1uMDexamethasone(DEX)respectivelyinDMEMand0.1%FBSfor16-24hours.
