Mammalian shRNA Tools for RNAi
- Constitutive Lentiviral
- Cre-regulated Lentiviral
- Tet-regulated Lentiviral
- Constitutive Retroviral
- RNA interference (RNAi) is an RNA-mediated gene silencing mechanism. As a tool in mammalian cell systems, silencing is achieved through the delivery of a double-stranded RNA (dsRNA) that matches the mRNA target sequence.
- The dsRNA can be delivered as an siRNA (short interfering RNA) via transfection, or shRNA (short hairpin RNA) via transfection or viral delivery of a plasmid.
- Below, browse our selection of empty vectors for cloning in shRNAs. Plasmids for constitutive expression as well as those that allow for conditional (Cre-lox) or inducible (Tet) expression are available. To find plasmids containing RNAi components, like Argonaute, please search our full site.
- Find resources for more information on shRNA design and delivery.
Constitutive Lentiviral shRNA
Lentiviral vectors for constitutive shRNA expression
|10878||pLKO.1 ‐ TRC cloning vector||shRNA expression; Recommended for cloning new shRNAs (contains a 1.9 kb stuffer region to easily visualize products from an AgeI/EcoRI restriction digest)||David Root|
|10879||pLKO.1 ‐ TRC control||Negative control vector containing non-hairpin insert||David Root|
|1864||pLKO.1 ‐ scrambled shRNA||Negative control vector containing scrambled shRNA||David Sabatini|
|11795||pLL3.7||Expresses shRNA under the mouse U6 promoter; A CMV-EGFP reporter cassette is included in the vector to monitor expression; Designed for inducing RNAi in a wide range of cell types, tissues and organisms||Luk Parijs|
|11619||pLB||Modified pLL3.7; Genetic elements known to prevent epigenetic silencing were added||Stephan Kissler|
|12247||pLVTHM||Expresses shRNA under the H1 promoter; shRNA or a H1-shRNA cassette from another vector (e.g. pSUPER) can be cloned into this vector; Contains a 5'LTR and should be packaged using the 2nd generation packaging system.||Didier Trono|
Cre-regulated Lentiviral shRNA
Lentiviral vectors for conditional (Cre-lox) shRNA expression
|11578||pSico||Cre addition causes EGFP to be recombined out of the construct, activating shRNA expression||Tyler Jacks|
|11586||pSico PGK puro||Cre addition causes the puromycin gene to be recombined out of the construct, activating shRNA expression||Tyler Jacks|
|11579||pSicoR||Cre addition causes both EGFP and shRNA to be recombined out of the construct, turning OFF shRNA expression||Tyler Jacks|
|12084||pSicoR PGK Puro||Cre addition causes both puromycin and shRNA to be recombined out of the construct, turning OFF shRNA expression||Tyler Jacks|
Tet-Regulated Lentiviral shRNA
Lentiviral vectors for Tet-regulated shRNA expression
|ID||Plasmid||Description||Tet on or off||PI|
|11643||pLVCT-tTR-KRAB||2nd generation; Transgene (CAG promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||On||Patrick Aebischer and Didier Trono|
|11779||pLVCT-rtTR-KRAB-2SM2||2nd generation; Transgene (CAG promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||Off||Patrick Aebischer and Didier Trono|
|11644||pLVET-tTR-KRAB||2nd generation; Transgene (hEF-1alpha promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||On||Patrick Aebischer and Didier Trono|
|11642||pLVPT-tTR-KRAB||2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||On||Patrick Aebischer and Didier Trono|
|11652||pLVPT-rtTR-KRAB-2SM2||2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||Off||Patrick Aebischer and Didier Trono|
|11651||pLVUT-tTR-KRAB||2nd generation; Transgene (hUbiquitin promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||On||Patrick Aebischer and Didier Trono|
|11648||pLVPRT-tTR-KRAB||2nd generation; Transgene (hPrion promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM)||On||Patrick Aebischer and Didier Trono|
Constitutive Retroviral shRNA
Retroviral vectors for constitutive shRNA expression
|8452||pMKO.1 puro||Retroviral vector for shRNA expression||Bob Weinberg|
|10675||pMKO.1 puro GFP shRNA||Negative control vector for pMKO.1 puro; Contains shRNA against GFP||William Hahn|
|10676||pMKO.1 GFP||Derivative of pMKO.1 puro with GFP instead of puromycin resistance gene||William Hahn|
- Read our detailed protocol for using the popular cloning vector pLKO.1.
- Trono Lab Lentivectors Toolbox
- Tyler Jacks Lab Protocols
Short hairpin RNA (shRNA): design, delivery, and assessment of gene knockdown. Moore CB, Guthrie EH, Huang MT, Taxman DJ. Methods Mol Biol. 2010. 629:141-58. PubMed.
A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Moffat J, Grueneberg DA, Yang X, Kim SY, Kloepfer AM, Hinkle G, Piqani B, Eisenhaure TM, Luo B, Grenier JK, Carpenter AE, Foo SY, Stewart SA, Stockwell BR, Hacohen N, Hahn WC, Lander ES, Sabatini DM, Root DE. Cell. 2006. 124:1283-1298. PubMed.
A versatile tool for conditional gene expression and knockdown. Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P. Nat Methods. 2006. Feb;3(2):109-16. PubMed.
In vivo RNA interference demonstrates a role for Nramp1 in modifying susceptibility to type 1 diabetes. Kissler S, Stern P, Takahashi K, Hunter K, Peterson LB, Wicker LS. Nat. Genet. 2006. Apr;38(4):479-83. PubMed.
Tuning silence: conditional systems for RNA interference. Wiznerowicz M, Szulc J, Trono D. Nat. Methods. 2006. Sep;3(9):682-8. PubMed.
Cre-lox-regulated conditional RNA interference from transgenes. Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L, Jaenisch R, Jacks T. PNAS. 2004. Jul 13; 101(28):10380-5. PubMed.
A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Rubinson DA, Dillon CP, Kwiatkowski AV, Sievers C, Yang L, Kopinja J, Rooney DL, Zhang M, Ihrig MM, McManus MT, Gertler FB, Scott ML, Van Parijs L. Nat. Genet. 2003. Mar; 33(3):401-6. PubMed.
Lentivirus-delivered stable gene silencing by RNAi in primary cells. Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD. RNA. 2003. 9(4):493-501. PubMed.