Description:
BMPResponsiveLuciferaseReporterHEK293CellLineisderivedfromhumanembyronickidney,andstablyexpressfireflyluciferasereportergeneunderthecontrolofthe Smad1/5/8responseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofBMPpathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
Bonemorphogenicprotein(BMP)isinvolvedinembryogenesis,developmentofmanyorgansystemsandadulttissuehomeostasis.Smads(Smad1/5/8)areactivatedduringthesignaltransductionandthenformacomplexwithSmad4,translocateintothenucleuswheretheyregulateexpressionoftranscriptionalfactorsandtranscriptionalcoactivators. DeficiencyinBMPproductionorfunctionalityusuallyleadstomarkeddefectsorseverehumandiseasesassociatedwithmostorgansystems. MonitoringtheBMPactivityisessentialtoresearchoftheBMPsignaling-associateddiseasesandconductdrugdiscovery.
Signosishasestablisheda BMPResponsiveLuciferaseReporterHEK293CellLinethathasbeenstablytransfectedwithpTA-BMP-luciferasereportervector,whichcontains4repeatsofBMPbindingsites,aminimalpromoterupstreamofthefireflyluciferasecodingregion.Therefore,thecelllinecanbeusedasareportersystemformonitoringtheactivationofBMPtriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisofSL-0051BMPreporteractivityinresponsetoBMPtreatment. Thecellswereseededona96-wellplateovernightincompletegrowthmedia. ThecellsthenweretreatedwithorwithoutBMPingrowthmediawith0.1%FBSfor16hours. TheBMPLuciferaseReporterHEK293CellLineexhibitsBMPdependentincreaseinluciferaseactivitywhencomparedtountreatedcells.
