Description:
GenMute™Reagentisanovelbiodegradablenon-LiposomalsiRNA/miRNA&DNAdeliverytool.WithourproprietarypHDependentConformationalChange(PDCC)technology,thebiodegradabletransfectionreagentwasformulatedbyadditionofpre-screenedhydrophobicgroupstoitssidechain,makingGenMute™Reagentaversatileandmostpowerfulgenedeliverytoolinthemarket.GenMute™ReagenthavebeenvalidatedtoeffectivelyandreproducIBLytransfectsinglesiRNA/miRNAorco-transfectDNA/siRNAorDNA/miRNAtovarietyofmammaliancells.

Size&content:
-GenMute™Reagent,1.0mL,sufficientfor~2,000reactionsbasedontransfecting5.0pmolsiRNAormiRNAmimicsin24-wellplate.
-GenMute™TransfectionBuffer(5x),formulatedformaximaltransfectionefficiency,8.0mLtomake40mLworkingsolution.

Applications:
-SinglesiRNAtransfectionorDNA/siRNAco-transfectiontovarietyofmammaliancells.
-SinglemiRNA,miRNAmimicsorinhibitorstransfectionorDNA/miRNAorDNA/miRNAmimicsco-transfectiontovarietyofmammaliancells.

Storage:
Storeat4°CforGenMute™ReagentandRTforGenMute™TransfectionBuffer(5x). Ifstoredproperly,theproductisstablefor12monthsorlonger.

Advantages:
-Excellentsilencingatlowconcentrationswithonly1.0nMsiRNA/miRNAmimics
-Minimizingnon-specificandofftargeteffectsinthecell
-Onetubereaction
-Bestforbroadmammaliancells
-Verylowcytotoxicity
-Veryaffordable

ComparisonsofSilencingEfficacyofGenMute™siRNA&DNATransfectionReagentwithBrandNameProducts

KnockdownefficacycomparisonofGenMute™TransfectionReagent(upperpanel)vs.Dharmafect™4siRNATransfectionReagent(middlepanel)andLipofectamine™2000(lowerpanel)onHEK293cells.siRNAtargetingrenillaluciferaseatdifferentfinalconcentrationsrangingfrom0.5to20nMwasco-transfectedwithrenillaluciferasegene(0.5µgofpRL-CMVDNAperwell)bytheabovethreetransfectionreagentspermanufacturers"protocolsintoHEK293cellsgrowingona24-wellplate.Renillaluciferaseactivitywasdetermined24hafterpostco-transfectionwithrenillaluciferasedeterminationsystem(Promega).Theluminescencewasmeasuredfrom5.0µloflysateduring10sintegrationwithaluminometer(BeckmanCoulterLD400).Luciferaseactivitywasexpressedaslightunitsintegratedover10s(RLU)andnormalizedpermgofcellproteinbyusingtheBCAassay.Theerrorsbarsrepresentstandarddeviationderivedfromtriplicateexperiments.Luciferase-silencingefficiencywascalculatedrelativetountreatedcells.WhileGenMute™andDharmafect™4reagentsdeliveredsignificantgenesilencingfrom1.0nMofrenillaluciferasesiRNA,lipofectamine™2000gavegoodknockdownonlyafter30nM(datanotshown).


ExcellentsilencingofendogenouslyexpressedKIF11(alsoknownasEg5)inHEK293cellswith0.5µlofGenMute™reagentand5.0pmolEg5siRNAperwellof24-wellplate.KIF11(alsoknownasEg5)encodesamotorproteinthatbelongstothekinesin-likeproteinfamilyinvolvedinchromosomepositioningandbipolarspindleformationduringcellmitosis.AreductioninKIF11levelscausesmitoticarrest.GenMute™reagenteffectivelydeliversEg5siRNA(final10nM,rightpanel)toHEK293cellsatonly0.5µlperwellof24-wellplatevs.negativecontrol(final10nM.leftpanel).Comparedwithnegativecontrol(leftpanel),phenotypeof"rounded-up"293cellswerevisualized24hoursposttransfection(rightpanel)withaNikonmicroscope.


GenMute™TransfectionReagentknockeddownendogenouslaminA/CgeneexpressioninHelacells.AsiRNAtargetinglaminA/Cgene(rightpanel)andashamsiRNA(leftpanel)wereintroducedintoHelacells(final1.0nM)byGenMute™TransfectionReagent.LaminA/Cgenesilencingwasmonitored24hposttransfectionbyimmunofluorescence.LaminA/CwasprobedwithamousemonoclonallaminA/CspecificantibodyfollowedbyadditionofFITC-conjugatedanti-mouseantibodies.QuantitativeanalysisshowedthatlaminsiRNAat1.0nMdeliveredbyGenMute™TransfectionReagentknockeddown94%endogenouslyexpressedlaminA/CinHelacells.

DataSheet&Protocol
-AProtocolforsiRNATransfectionandDNA/siRNACo-Tranafection
-AProtocolformiRNAMimicsTransfectionandDNA/miRNACo-Transfection
-AProtocolforDNATransfectiontoMammalianCells

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