Purpose(Empty Backbone) 2nd gen lentivector expressing shRNA from H1 promoter and GFP. For direct cloning of shRNA or sub-cloning of an H1-shRNA cassette from pSUPER. Dox-controllable if used with tTRKRAB expressing plasmid
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||12247||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 11087
Vector typeMammalian Expression, Lentiviral, RNAi, Cre/Lox
Growth in Bacteria
Growth instructionsUse Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer See map (Common Sequencing Primers)
Terms and Licenses
Articles Citing this Plasmid
pLVTHM allows for direct cloning of shRNAs into the lentiviral vectors and replaces the old pLVTH system from Trono lab.
pLVTHM is similar to pLVTH, but contains a 3bp substitution that generates a unique MluI site for direct cloning of an shRNA into MluI-ClaI.
Please note that there is an additional ClaI site in this vector that is blocked by Dam methylation (this site does not appear in the depositor's full sequence). The plasmid needs to be grown in a dam(-) bacterial strain in order to use ClaI for cloning.
Note that ClaI has lower salt concentration requirements than MluI. One way to handle this is to digest with ClaI for 45 minutes, followed by addition of diluted MluI buffer and MluI, then incubation for 90 more minutes. Be sure not to use a large amount of vector (1.7 ug to 2.5 ug is known to work).
Also, if your shRNA is already in pSUPER (or another plasmid under the control of the PolIII promoter) you may use the EcoRI-ClaI sites to replace the H1 promoter in pLVTHM with the H1-shRNA cassette from pSUPER (or another plasmid).
The packaging plasmid for Trono lab lentiviral vectors is also available at Addgene http://www.addgene.org/rnaitools
Please note that the full sequence for this plasmid is approximated and not fully verified. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussions 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:pLVTHM was a gift from Didier Trono (Addgene plasmid # 12247 ; http://n2t.net/addgene:12247 ; RRID:Addgene_12247)
For your References section:Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. Wiznerowicz M, Trono D. J Virol. 2003 Aug . 77(16):8957-61. 10.1128/JVI.77.16.8957-8951.2003 PubMed 12885912