Addgene - pLVTHM Plasmid Data

Gene/insert name:
None
Vector backbone:
pLVTH
(Search Vector Database)
Vector type:
Mammalian Expression, Lentiviral, RNAi, Cre/Lox
Backbone size w/o insert:
11087
5' sequencing primer:
See map List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
Stbl3
Growth temperature (℃):
37
Growth instructions:
Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
High or low copy:
High Copy
Sequence: View sequences (2)
Map:
View map
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Didier Trono
Terms and Licenses:
MTA

Comments:

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. 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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. Wiznerowicz et al (J Virol. 2003 Aug . 77(16):8957-61. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 12247" in your Materials and Methods section.