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Signosis/TCF/LEF Luciferase Reporter HEK293 Stable Cell Line/SL-0015 NP/1 Ea
Description:
TCF/LEFResponsiveLuciferaseReporterHEK293StableCellLineisderivedfromhumanembryonickidney,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheTCF/LEF responseelement. Thiscelllineisanidealcellularmodelformonitoringtheactivation ofWnt/b-catenin ReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
TheWntsignalingpathwayisimportanttobothembryonicdevelopmentandtumOrigenesis.β-Catenin,thecentralcomponentofthepathway,functionsasaco-factoroftheTCF/LEFfamilyoftranscriptionfactors. Together,theyactivatetranscriptionofWnttargetgenesbybindingtothepromotersofdownstreamtargetgenesinvolvedincellproliferation,survival,andmigration.
SignosishasestablishedaTCF/LEFluciferasereporterstablecelllinethathasbeenstablytransfectedwithpTA-TCF/LEF-luciferasereportervector,whichcontains6repeatsofTCF/LEFbindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion. Therefore,thecelllinecanbeusedasareportersystemformonitoringtheactivationofβ-catenintriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisofTCF/LEFLuciferaseReporterHEK293StableCellLine.HEK293cellsstablyexpressingTCF/LEFluciferasereporterweretreatedwith10mMLiClor40ng/mLWnt3atoactivateb-cateninsignaling. 16hourstreatmentofLiClinduced100foldluciferaseactivitiesandthecombinedtreatmentofLiClandWnt3arobustlyinducedtheluciferaseactivitiesover500fold.
