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Plasmid 17619: EF.CMV.RFP
Comments: 

In a SIN LV containing the elongation factor 1alpha promoter, we included a second promoter from cytomegalovirus that drives expression of RFP as a reporter. Dual-promoter LVs can coexpress multiple transgenes efficiently in a single target cell.

Recombinant LVs were produced by transient transfection of the transducing vector into 293T cells with two packaging vectors: pMD.G, a plasmid expressing the VSV-G envelope gene, and pCMVDeltaR8.91, a plasmid expressing the HIV-1 gag/pol, tat, and rev genes.

There is an a->g mutation in this plasmid that abolishes the HpaI site. A 10bp insertion in Addgene sequence corresponding to position 6893, 21bp insertion in Addgene sequence at position 7405, and 11bp deletion in Addgene sequence at position 7544 do not alter function of the plasmid or RFP expression.

The uploaded author's map shows an example with GFP cloned downstream of the EF-1a promoter. The vector supplied by Addgene does not contain the GFP.

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

Article: Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells. Yu et al (Mol Ther. 2003 Jun . 7(6):827-38. 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 17619" in your Materials and Methods section.

Price: US $65

Available to academic and non-profits only