Description:
ERResponsiveLuciferaseReporterT47DStableCellLineisderivedfromhumanbreastcancer,andstablyexpressfireflyluciferasereportergeneunderthecontroloftheERresponseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofEstrogenReceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
Estrogenreceptor(ER)belongstonuclearreceptorfamilyandplaysawidespreadroleinhumanphysiologyandinthedevelopmentorprogressionofnumerousdiseases. Inresponsetoestrogenstimulation,estrogenboundreceptorinthenucleusdimerizesandbindstospecificresponseelementsknownasestrogenresponseelements(EREs)locatedinthepromotersoftargetgenesandregulatestheirgeneexpression.SignosishasestablishedtheT47DERluciferasereporterstablecellline,inwhichtheEREandreporterluciferasegeneareconsistentlyexpressedinthecelllinetofacilitatethescreeningandstudy.Thisstablecelllinecanprovideasensitive,responsive,andrapidinvitrosystemtodetectandmeasuresubstanceswithpotential(anti-)estrogenicactivity.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
T47D/ER-luccellsweretreatedwithvariousconcentrationsinresponseto17β-EstrADIol.
