Pre-madelentiviralparticlesbicistronicallyexpresstwoindividualproteins:nuclearpermeableCRE recombinaseandtheGFPMarkerunderthesameenahncedEf1apromoter.CREcontainsaNLS(nuclearlocalizationsignal) atitsN-term.This nuclearpermeant CRE increasestherecombinationeventsinvivo.APuromycinmarkerwasexpressedunderRSVpromoter.ParticlesallowstosortorselecttransducedcellsforlongtermexpressionviaGFPsignalorviaPuromycinantibioticselection.
CRE protein,frombacteriophageP1,catalyzesrecombinationbetween34basepairtargetsequences, namedloxsites.CRElentiviralparticles can catalyze thejointingeventInVivo inthepresenceof loxsites.PurifiedCREenzymecanjointindividualplasmidscontainingloxsites.
CREandGFPareconstitutivelyexpressedinhighlevel,indenpendentofcelltypes.GFPsignalprovidesaconvenient,realtimemonitoringthe particlesperformance. Seeproductmanualfordetails(.pdf). Amount: 200ul/pervial,at1x107IFU/mlinDMEMmediumwith10%FBSand10xpolybrene. Cat#:LVP526
