Bicistronic Lentiviral Vector Systems
Bicistronic Lentiviral Vector Systems
Brand: Takara Bio.
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Bicistronic Lentiviral Vector Systems
Lenti-X bicistronic lentiviral vectors and expression systems allow you to deliver and stably express your gene of interest, along with a coexpressed bright fluorescent protein or an antibiotic selection marker, in virtually any mammalian cell type. The coexpressed proteins are produced from a single mRNA transcript, so you can be assured that your protein of interest is present in cells in which the detectable marker is also present.
The Lenti-X Bicistronic Expression System is available in three different formats with expression vectors containing Neor, Hygr, or Puror markers. Alternatively, you can coexpress your gene of interest with our Living Colors mCherry, tdTomato, and ZsGreen1 fluorescent proteins using the pLVX-IRES-mCherry, pLVX-IRES-tdTomato, and pLVX-IRES-ZsGreen1 Vectors. These are supremely bright fluorescent proteins that are excellent reporters for gene expression or for measuring the efficiency of transduction or transfection. mCherry is a very popular mutant derived by directed mutagenesis of mRFP1, a monomeric mutant of DsRed.
Simultaneous expression of two proteins—how does it work?
The key element of these bicistronic lentiviral vectors is an optimized internal ribosome entry site (IRES) that permits your gene of interest and a second detectable protein to be coexpressed from a single mRNA transcript. While translation initiation of eukaryotic mRNAs occurs almost exclusively at the 5' cap, the IRES lures ribosomes to begin translation at a second, internal location. The result is that two proteins are expressed simultaneously from a single bicistronic mRNA transcript.
Overview
- Lenti-X packaging single shots are provided with each expression system
- Track expression of your protein using fluorescence microscopy
- Expression of your protein is assured in antibiotic-resistant clones
- Bright fluorescent protein coexpression—learn more from our fluorescent protein quick guide
- Monitor the expression of your lentiviral-delivered genes with fluorescent tags and antibiotic selection markers
Lentiviral vector design
Lentiviral vector design. Clontech's lentiviral vectors contain sequence elements that facilitate lentiviral packaging and boost transgene expression. The Lenti-X Expression System (pLVX-Puro Vector) is used for constitutive expression of your transgene. The Lenti-X Bicistronic Expression Systems allow your transgene and a selectable marker to be coexpressed from a single mRNA transcript.
High-titer lentivirus production
High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.
4th generation lentiviral packaging system
Fourth-generation lentiviral packaging system. A lentiviral vector and the Lenti-X HTX Packaging Mix are cotransfected into 293T cells. High titer lentiviral supernatants are ready for use 48 hr after transfection.
Lentiviral vectors with fluorescent proteins
Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.
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Lentiviral vector design
Lentiviral vector design. Clontech's lentiviral vectors contain sequence elements that facilitate lentiviral packaging and boost transgene expression. The Lenti-X Expression System (pLVX-Puro Vector) is used for constitutive expression of your transgene. The Lenti-X Bicistronic Expression Systems allow your transgene and a selectable marker to be coexpressed from a single mRNA transcript.
High-titer lentivirus production
High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.
4th generation lentiviral packaging system
Fourth-generation lentiviral packaging system. A lentiviral vector and the Lenti-X HTX Packaging Mix are cotransfected into 293T cells. High titer lentiviral supernatants are ready for use 48 hr after transfection.
Lentiviral vectors with fluorescent proteins
Lentiviral vectors with fluorescent proteins. Lenti-X vectors contain sequence elements that facilitate lentiviral packaging and/or boost transgene expression, including the LTRs, packaging signal (Ψ), Rev response element (RRE), and central polypurine tract/central termination sequence (cPPT/CTS) from HIV-1; and the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). Vectors can express your protein fused at its N- or C- terminus to either a green (AcGFP1) or red (DsRed-Monomer) fluorescent protein tag, or coexpress it as a separate protein along with ZsGreen1 (shown), mCherry, or tdTomato.
