>
产品中心 >
Stable_cell_line >
Signosis/CREB Luciferase Reporter NIH 3T3 Stable Cell Line/SL-0031-NP/1 Ea
Description:
CREBResponsiveLuciferaseReporterNIH/3T3StableCellLineisderivedfrommousefibroblast,andstablyexpressfireflyluciferasereportergeneunderthecontrolofCREBresponseelement. ThiscelllineisanidealcellularmodelformonitoringtheactivationofcAMP,PKA,CaMKSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
CyclicAMPresponseelement(CRE)-bindingprotein(CREB)isatranscriptionfactorthatregulatesandrespondstodiversecellularresponses,rangingfromproliferation,survival,differentiation,stressresponses,andneuronalactivity. Thesecellularsignalsleadtoupstreamkinaseactivation,suchasproteinkinaseA(PKA),pp90ribosomalS6kinase(pp90RSK),andCa2+/calmodulin-dependentproteinkinases(CaMKs),andthesekinasesinturnphosphorylateCREBtoinduceCREBactivity. CREBincreasesthetranscriptionofgenesthatcontaincAMPresponsiveelements.
SignosishasdevelopedCREBluciferasereporterstablecelllinebyco-transfectingCREBluciferasereportervectorandhygromycinexpressionvector. Thehygromycinresistantclonesweresubsequentlyscreenedforforskolin-inducedluciferaseactivity. ThecelllinecanbeusedasareportersystemformonitoringtheactivationofCREBtriggeredbystimulitreatment,suchasforskolinandgeneoverexpressionandgeneknockdown.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisofSL-0031CREBreporteractivityinresponsetoforskolintreatment. Thecellswereseededona96-wellplateforovernightwithDMEMincluding10%FBS.Thecellsthenweretreatedwithorwithout10uMforkoslininDMEMand0.1%FBSfor16hours. Forskolininducedover10-foldincreaseinluciferaseactivitycomparedtountreatedcells.
