GentargetprovidesdifferentsetsofpremadelentivirialparticlesforhumanormouseiPSgenes(seeiPSparticleproductpage).
ThispremadelentiviralparticlesisthesinglevectormethodforiPScellgeneration.FourmouseiPSfactor,c-Myc,Klf4,Oct4andSox2(MKOS)wereclonedunderthesameTetracyclineinducIBLeCMVpromoter.All4factorswereclonedinframebutseparatedbyF2Asignalfragment,whichresultintheexpressionoffourindividualfactorvia“ribosomalskipping”mechanism.Inaddition,theparticlescontainsaRFP-blasticidindualselectionMarkerunderRSVpromoter,whichprovidesaconvenience mothodtomonitortheiPSfactorexpressionefficiency,andtosortthepositveipsexpressioncells.
ThefouriPSfactorareexpressedunderanoptionalinducibleCMVpromoter.ThepremadeparticlecanbeusedasconstitutiveexpressionoffouriPSfactors,butoptionally,itcanbeusedasantetracyclineinducibleexpressionofiPSfactors.Tousedasinducibleexpression,therepressorprotein(TetR)mustbepresentinadvancetostoptheexpression,thentheexpressionofiPSfactorsonlyoccurswhentheTetracyclineorDoxwasaddedintothecellculture.PleaseclickheretoseemoredetailsabouttheOptionalinduciblelentiviralexpression.
Thisparticlewasproducedbyco-transfectionofthechainediPSinduciblelentivectorwithapackagingmix(Cat#HT-pack)intoa293Tpackagingcells(cat#TLV-C).Thepre-madelentiviralparticlesareVSV-Gpseudotypedvirus,packagedin10%FBSmedium,andsuppliedas500ul/pervialat~1x106IFU/ml.Seeproductmanual(.pdf)fordetails.
Amounts:mMKOS: 0.5ml/tubex 1tubes, at1x106IFU/ml
Cat#:LVP381
