We have developed different lentiviral vector systems to generate cell lines for gene expression or overexpression.
Lentiviral Vectors and Viruses
The pLV-EF1a-GOI-IRES-marker vector system
The selection markers are expressed from IRES to favor the transduced cells with higher copy numbers of lentiviral genome.
The level of gene expression from IRES is generally around 7-10 times lower than the same gene expressed directly from the promoter before IRES. Therefore, the antibiotic selection will favor the transduced cells with higher copy numbers. The cells transduced with for example 1-5 copies of viruses may not survive after selection, especially for the cells with lower transduction efficiency such as primary cell, blood cells, and rodent cells. This vector system is good for overexpression but transduction efficiency will be important and higher MOIs, e.g. 10-50, transduction is recommended.
The dual promoter vector system
The gene of interest (GOI) is expressed from the human EF1a promoter and the antibiotic selection marker is expressed from the mouse PGK promoter. The dual promoter vector system gives the user more flexibility to generate stable cell lines with a broader range of gene expression by using low, medium, and high MOIs.
The pLV-EF1a-marker-IRES-GOI vector system
The GOI is expressed from IRES to closed to the endogenous level
The pLV-EF1a-2A vector system
The pLV-EF1a-2A vector system allows users to turn the lentiviral vector into a polycistronic expression cassette in which the multiple genes are joined by 2A peptides and expressed from a single open reading frame.
