11/12/2016

Addgene:MammalianRNAiTools

MammalianshRNAToolsforRNAi
ConstitutiveLentiviral | CreregulatedLentiviral | TetregulatedLentiviral | ConstitutiveRetroviral | Resources

Background

AdditionalResources:

RNAinterference(RNAi)isanRNAmediatedgenesilencingmechanism.Asatoolinmammaliancell
systems,silencingisachievedthroughthedeliveryofadoublestrandedRNA(dsRNA)thatmatchesthe
mRNAtargetsequence.

Humanandmousepooled
shRNAlibraries

ThedsRNAcanbedeliveredasansiRNA(shortinterferingRNA)viatransfection,orshRNA(shorthairpin
RNA)viatransfectionorviraldeliveryofaplasmid.

Lentiviralpackagingand
expressionplasmids

Below,browseourselectionofemptyvectorsforcloninginshRNAs.Plasmidsforconstitutiveexpressionas
wellasthosethatallowforconditional(Crelox)orinducible(Tet)expressionareavailable.Tofindplasmids
containingRNAicomponents,likeArgonaute,pleasesearchourfullsite.
FindresourcesformoreinformationonshRNAdesignanddelivery.

ConstitutiveLentiviralshRNA
LentiviralvectorsforconstitutiveshRNAexpression
ID

Plasmid

Description

PI

10878

pLKO.1TRC
cloningvector

shRNAexpressionRecommendedforcloningnewshRNAs(containsa1.9kbstuffer
regiontoeasilyvisualizeproductsfromanAgeI/EcoRIrestrictiondigest)

David
Root

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10879

pLKO.1TRC
control

Negativecontrolvectorcontainingnonhairpininsert

David
Root

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1864

pLKO.1
scrambled
shRNA

NegativecontrolvectorcontainingscrambledshRNA

David
Sabatini

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11795

pLL3.7

ExpressesshRNAunderthemouseU6promoterACMVEGFPreportercassetteis
includedinthevectortomonitorexpressionDesignedforinducingRNAiinawide
rangeofcelltypes,tissuesandorganisms

Luk
Parijs

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11619

pLB

ModifiedpLL3.7Geneticelementsknowntopreventepigeneticsilencingwereadded

Stephan
Kissler

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12247

pLVTHM

ExpressesshRNAundertheH1promotershRNAoraH1shRNAcassettefrom
anothervector(e.g.pSUPER)canbeclonedintothisvectorContainsa5'LTRand
shouldbepackagedusingthe2ndgenerationpackagingsystem.

Didier
Trono

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CreregulatedLentiviralshRNA
Lentiviralvectorsforconditional(Crelox)shRNAexpression
ID

Plasmid

Description

PI

11578

pSico

CreadditioncausesEGFPtoberecombinedoutoftheconstruct,activatingshRNA
expression

Tyler
Jacks

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11586

pSicoPGK
puro

Creadditioncausesthepuromycingenetoberecombinedoutoftheconstruct,
activatingshRNAexpression

Tyler
Jacks

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11579

pSicoR

CreadditioncausesbothEGFPandshRNAtoberecombinedoutoftheconstruct,
turningOFFshRNAexpression

Tyler
Jacks

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12084

pSicoRPGK

CreadditioncausesbothpuromycinandshRNAtoberecombinedoutofthe

Tyler

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http://www.addgene.org/mammalianrnai/

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Addgene:MammalianRNAiTools

Puro

construct,turningOFFshRNAexpression

Jacks

TetRegulatedLentiviralshRNA
LentiviralvectorsforTetregulatedshRNAexpression
Teton
oroff

ID

Plasmid

Description

PI

11643

pLVCTtTR
KRAB

2ndgenerationTransgene(CAGpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

On

Patrick
Aebischerand
DidierTrono

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11779

pLVCT
rtTRKRAB
2SM2

2ndgenerationTransgene(CAGpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

Off

Patrick
Aebischerand
DidierTrono

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11644

pLVETtTR
KRAB

2ndgenerationTransgene(hEF1alphapromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

On

Patrick
Aebischerand
DidierTrono

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11642

pLVPTtTR
KRAB

2ndgenerationTransgene(hPGKpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

On

Patrick
Aebischerand
DidierTrono

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11652

pLVPT
rtTRKRAB
2SM2

2ndgenerationTransgene(hPGKpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

Off

Patrick
Aebischerand
DidierTrono

AddtoCart

11651

pLVUTtTR
KRAB

2ndgenerationTransgene(hUbiquitinpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

On

Patrick
Aebischerand
DidierTrono

AddtoCart

11648

pLVPRT
tTRKRAB

2ndgenerationTransgene(hPrionpromoter)ORshRNA(H1
promoterwhensubclonedfrompLVTHM)

On

Patrick
Aebischerand
DidierTrono

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ConstitutiveRetroviralshRNA
RetroviralvectorsforconstitutiveshRNAexpression
ID

Plasmid

Description

PI

8452

pMKO.1puro

RetroviralvectorforshRNAexpression

Bob
Weinberg

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10675

pMKO.1puro
GFPshRNA

NegativecontrolvectorforpMKO.1puroContainsshRNAagainstGFP

William
Hahn

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10676

pMKO.1GFP

DerivativeofpMKO.1purowithGFPinsteadofpuromycinresistancegene

William
Hahn

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AdditionalResources
WebReferences
ReadourdetailedprotocolforusingthepopularcloningvectorpLKO.1.
TronoLabLentivectorsToolbox
TylerJacksLabProtocols

Publications
ShorthairpinRNA(shRNA):design,delivery,andassessmentofgeneknockdown.MooreCB,GuthrieEH,HuangMT,TaxmanDJ.Methods
MolBiol.2010.629:14158.PubMed.

http://www.addgene.org/mammalianrnai/

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AlentiviralRNAilibraryforhumanandmousegenesappliedtoanarrayedviralhighcontentscreen.MoffatJ,GruenebergDA,YangX,Kim
SY,KloepferAM,HinkleG,PiqaniB,EisenhaureTM,LuoB,GrenierJK,CarpenterAE,FooSY,StewartSA,StockwellBR,HacohenN,Hahn
WC,LanderES,SabatiniDM,RootDE.Cell.2006.124:12831298.PubMed.
Aversatiletoolforconditionalgeneexpressionandknockdown.SzulcJ,WiznerowiczM,SauvainMO,TronoD,AebischerP.NatMethods.
2006.Feb3(2):10916.PubMed.
InvivoRNAinterferencedemonstratesaroleforNramp1inmodifyingsusceptibilitytotype1diabetes.KisslerS,SternP,TakahashiK,Hunter
K,PetersonLB,WickerLS.Nat.Genet.2006.Apr38(4):47983.PubMed.
Tuningsilence:conditionalsystemsforRNAinterference.WiznerowiczM,SzulcJ,TronoD.Nat.Methods.2006.Sep3(9):6828.PubMed.
CreloxregulatedconditionalRNAinterferencefromtransgenes.VenturaA,MeissnerA,DillonCP,McManusM,SharpPA,VanParijsL,
JaenischR,JacksT.PNAS.2004.Jul13101(28):103805.PubMed.
Alentivirusbasedsystemtofunctionallysilencegenesinprimarymammaliancells,stemcellsandtransgenicmicebyRNAinterference.
RubinsonDA,DillonCP,KwiatkowskiAV,SieversC,YangL,KopinjaJ,RooneyDL,ZhangM,IhrigMM,McManusMT,GertlerFB,ScottML,
VanParijsL.Nat.Genet.2003.Mar33(3):4016.PubMed.
LentivirusdeliveredstablegenesilencingbyRNAiinprimarycells.StewartSA,DykxhoornDM,PalliserD,MizunoH,YuEY,AnDS,Sabatini
DM,ChenIS,HahnWC,SharpPA,WeinbergRA,NovinaCD.RNA.2003.9(4):493501.PubMed.

http://www.addgene.org/mammalianrnai/

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