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shRNA-test.png Mammalian shRNA Tools for RNAi


Background

  • 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

   ID Plasmid Description PI   
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 Add to Cart
10879 pLKO.1 ‐ TRC control Negative control vector containing non-hairpin insert David Root Add to Cart
1864 pLKO.1 ‐ scrambled shRNA Negative control vector containing scrambled shRNA David Sabatini Add to Cart
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 Add to Cart
11619 pLB Modified pLL3.7; Genetic elements known to prevent epigenetic silencing were added Stephan Kissler Add to Cart
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 Add to Cart

Cre-regulated Lentiviral shRNA

Lentiviral vectors for conditional (Cre-lox) shRNA expression

   ID Plasmid Description PI   
11578 pSico Cre addition causes EGFP to be recombined out of the construct, activating shRNA expression Tyler Jacks Add to Cart
11586 pSico PGK puro Cre addition causes the puromycin gene to be recombined out of the construct, activating shRNA expression Tyler Jacks Add to Cart
11579 pSicoR Cre addition causes both EGFP and shRNA to be recombined out of the construct, turning OFF shRNA expression Tyler Jacks Add to Cart
12084 pSicoR PGK Puro Cre addition causes both puromycin and shRNA to be recombined out of the construct, turning OFF shRNA expression Tyler Jacks Add to Cart

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 Add to Cart
11779 pLVCT-rtTR-KRAB-2SM2 2nd generation; Transgene (CAG promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) Off Patrick Aebischer and Didier Trono Add to Cart
11644 pLVET-tTR-KRAB 2nd generation; Transgene (hEF-1alpha promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) On Patrick Aebischer and Didier Trono Add to Cart
11642 pLVPT-tTR-KRAB 2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) On Patrick Aebischer and Didier Trono Add to Cart
11652 pLVPT-rtTR-KRAB-2SM2 2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) Off Patrick Aebischer and Didier Trono Add to Cart
11651 pLVUT-tTR-KRAB 2nd generation; Transgene (hUbiquitin promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) On Patrick Aebischer and Didier Trono Add to Cart
11648 pLVPRT-tTR-KRAB 2nd generation; Transgene (hPrion promoter) ‐ OR ‐ shRNA (H1 promoter when subcloned from pLVTHM) On Patrick Aebischer and Didier Trono Add to Cart

Constitutive Retroviral shRNA

Retroviral vectors for constitutive shRNA expression

   ID Plasmid Description PI   
8452 pMKO.1 puro Retroviral vector for shRNA expression Bob Weinberg Add to Cart
10675 pMKO.1 puro GFP shRNA Negative control vector for pMKO.1 puro; Contains shRNA against GFP William Hahn Add to Cart
10676 pMKO.1 GFP Derivative of pMKO.1 puro with GFP instead of puromycin resistance gene William Hahn Add to Cart

Additional Resources

Web References

Publications

  • 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.