Signosis/HIF Luciferase Reporter Neuro2a Stable Cell Line/SL0027FP/1 Ea

Description:

HIFresponsiveLuciferaseReporterNeuro2a StableCellLineisderivedfrommouseneuroblastoma,andstablyexpressfireflyluciferasereportergeneunderthecontrolof HIFresponseelement.ThiscelllineisanidealcellularmodelformonitoringtheactivationofHypoxiareceptorSignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.

Principle:

Hypoxia-InducibleFactor(HIF)-1isadimericproteincomplexthatplaysacentralroleintheresponsetolowoxygenconcentrations,orhypoxia,and isacrucialphysiologicalregulatorofhomeostasis,vascularization,andanaerobicmetabolism.HIFisatranscriptionfactorregulatinggeneexpressionbybindingtotheirDNArecognitionsiteonthetargetgenes.Furthermore,HIF-1hasbeenwidelystudiedbecauseofitsperceivedtherapeuticpotential. HIF-1allowssurvivalandproliferationofcancerouscellsduetoitsangiogenicproperties,andtheinhibitionpotentiallycouldpreventthespreadofcancer.WithagrowingunderstandingoftheHIF-1pathway,ithasbecomeanattractivegoaltoanalyzetheinhibitionandstimulationofHIFtranscriptionalactivityviasmallmolecules. 

Signosishasdeveloped HIFLuciferaseReporterStableCellLinebytransfectionofHIFluciferasereportervectoralongwithhygromycinexpressionvectorfollowedbyhygromycinselection.ThehygromycinresistantclonesweresubsequentlyscreenedforCoCl2orDMOG-inducedluciferaseactivity.

 

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