|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||11777||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 11627
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Growth instructionsUse Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Copy numberHigh Copy
Gene/Insert namemPGK, GFP, rtTR-KRAB, Tet-off
- Cloning method Restriction Enzyme
- 5′ sequencing primer See map (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
Article Citing this Plasmid
Transgenes can be expressed from any RNA Pol II promoter as part of bicistronic unit comprising the KRAB-based repressor; tetO sequences are inserted into the vector LTR. Tet-on and Tet-off versions rely on repressors that bind in the absence or the presence of doxycycline, respectively. Addition of Pol III promoter-small hairpin RNA cassette allows for drug-controllable RNA interference (Tet-on shRNA).
Cloning shRNA into pLVET, pLVCT, and pLVPT vectors: first clone shRNA into pLVTHM downstream of the tetO-H1 region. Then cut pLVTHM containing your shRNA with MscI-FspI and clone the insert containing the 3'LTR to target plasmid opened with MscI-FspI. FspI cuts into AmpR. This means for inverted clones AmpR will not be restored; after selection you will be left with clones with the shRNA cassette and functional AmpR.
pLVTHM and packaging plasmid for this system are also available at Addgene http://www.addgene.org/rnaitools Please visit Trono lab website http://tronolab.epfl.ch to see frequently asked questions on cloning strategies and packaging. You may also visit LentiWeb http://www.lentiweb.com for discussion on cloning strategies and protocols.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pLVPT-rtTR-KRAB was a gift from Patrick Aebischer & Didier Trono (Addgene plasmid # 11777 ; http://n2t.net/addgene:11777 ; RRID:Addgene_11777)
For your References section: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. 10.1038/nmeth846 PubMed 16432520