Signosis/GR Luciferase Reporter Hela Stable Cell Line/SL0021NP/1 Ea

Description:

GRResponsiveLuciferaseReporterHelaStableCellLineisderivedfromhumancervicalcancer,andstablyexpressfireflyluciferasereportergeneunderthecontrolofGRresponseelement.Thiscelllineisanidealcellularmodelformonitoringtheactivation ofGlucocorticoidSignalingReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.

Principle:

TheGlucocorticoidreceptor(GR)isamajorplayerindevelopment,metabolismandimmuneresponse. Whenactivatedbystimuli,GRtranslocatesfromthecytoplasmintothenucleus,andthenbindsaDNArecognitionsitetoregulategeneexpression. Inaddition,activatedGRcantransrepressothertranscriptionfactorsthathavebeenmisregulatedincancerandotherdiseases. 

SignosishasestablishedaGRluciferasereporterstablecellline,bytransfectionofaGRfireflyluciferasereportervectoralongwitheitherG418orhygromycinexpressionvectorfollowedbyG418orhygromycinselection,respectively.Theantibioticresistantclonesweresubsequentlyscreenedfordexamethasone(DEX)-inducedluciferaseactivity.Theclonewiththehighestfoldinductionwasselectedandexpandedforproduction.

 

PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation.

 

Data:

 AnalysisofGRLuciferaseReporterHelacellline. Thecellswereseededona96-wellplateforovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout2uMDexamethasone(DEX)respectivelyinDMEMand0.1%FBSfor16hours. Closeto100foldincreaseinluciferaseactivitywasdetectedwhencomparedtountreatedcells.