Purpose(Empty Backbone) 3rd gen lentiviral backbone for cloning and expression of new shRNA sequences. Uses puromycin for selection.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||8453||Plasmid sent as bacteria in agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 7032
Vector typeMammalian Expression, Lentiviral, RNAi
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ cloning site AgeI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer LKO.1 5' (Common Sequencing Primers)
Empty lentiviral vector for siRNA expression; replaces Lentihair; see author's map. The 293T cell line can be obtained from the Weinberg lab or GenHunter http://genhunter.com/products/aptag-3/index.html
Note that Addgene's quality control sequence shows that there are about 25 nucleotides between the AgeI and EcoRI sites (the depositor sequences is not as accurate in this region).
For packaging, please use pCMV-dR8.2 dvpr (Addgene plasmid #8455) and pCMV-VSVG (Addgene plasmid #8454). For the official vector of The RNAi Consortium and a plasmid map, please see plasmid #10878.
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:pLKO.1 puro was a gift from Bob Weinberg (Addgene plasmid # 8453)
For your References section: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 Apr;9(4):493-501. 10.1261/rna.2192803 PubMed 12649500