Overview

Lentiviruses are a group of retroviruses that can infect and deliver genetic material into both dividing and non-dividing cells.. In therapy and genetic engineering, recombinant lentiviral system is commonly used to deliver genes of interest (GOI) into cells. Upon integration into the host genome, GOI are permanently expressed. The lentiviral transgene plasmid (pTransgene) is a key component of this recombinant system: this plasmid encodes the GOI to be transferred and contains lentiviral DNA sequences required for the integration of the transgene into the host genome following viral transduction. To be safely manipulated, pTransgenes are designed to be replication incompetent.

To produce lentiviral particles, enable to efficiently deliver the GOI into cells of interest, we have developed an 3rd generation Easy lentiviral pTransgene plasmid to be co-transfected into packaging cells along with packaging lentiviral plasmids encoding the Gag, Pol, Rev, VSV-G viral components, and with the transfection reagents (e.g. FectoVIR®-LV).

 

Genetic Features :

5’and 3’ lentiviral arms : These two sequences contain important structural and regulatory elements necessary for the transgene transcription and integration into the host cell’s genome. These elements include the CMV viral promoter region, modified long terminal repeat (LTR) sequences, the cPPT sequence, the Psi sequence, the RRE sequence and a PolyA signal.

eZ-Ori : The e-Zyvec® Origin of replication is ahigh copy number origin of replication (pMB1 Ori) optimized by e-Zyvec for optimal assembly and plasmid amplification yield in E.coli.

GOI : The GOI (gene of interest) is the sequence that you wish to see expressed in your target cells.
KanR : The prokaryotic kanamycin resistance cassette comprises a promoter, the nptII gene and a hairpin terminator, to provide antibiotic resistance to bacteria expressing it.

Kozak : Is a sequence of nucleotides in eukaryotic mRNA that is recognized and used by the ribosome as the start site for translation.

Promoter : The promoter is the DNA sequence that will drive the expression of your gene of interest. The expression level of your gene of interest is cell-type and promoter-dependent. We offer a variety of promoters to fit your needs.

PuroR : The Puromycin resistance gene (Pac gene) represents in the Polyplus easy lentiviral transgene the eukaryotic selectable marker that will allow you to select transduced cells. In this easy plasmid, its expression is driven by the constitutive Simian Virus 40 promoter.

WPRE : the”Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element.” significatntly enhances mRNA stability and nuclear export, thus increasing the expression level of transgenes.

If you require a more specific set of features for your plasmid, we provide a fully customizable plasmid construction service, do not hesitate to contact us regarding your projects.

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