Takara Lenti-X Bicistronic Expression System (Puro) \Each

The Lenti-X Bicistronic Expression Systems enable you to produce high-titer recombinant lentivirus for the purpose of transferring genes into dividing or non-dividing mammalian cells. The pLVX-IRES-Puro Vector contains an internal ribosome entry site (IRES) followed by a drug resistance marker, SO that your gene of interest and puromycin resistance are simultaneously coexpressed from a single mRNA transcript. The Lenti-X packaging system is a proprietary combination of plasmids that collectively expresses all the viral proteins necessary for assembling the vector into infectious virions. Lenti-X Packaging Single Shots, which contain lyophilized Lenti-X packaging plasmids and transfection reagent, are used for efficient introduction of the packaging plasmids and the pLVX-IRES-Puro Vector into Lenti-X 293T Cells (Cat. No. 632180; not included) to produce lentivirus. These optimized reagents allow the transfected Lenti-X 293T cultures to produce high titers of VSV-g pseudotyped lentivirus that is both safe and replication-incompetent, and able to transduce virtually any cell type. The Lenti-X GoStixcan be used to determine whether virus production is within a usable range or for selecting the best time to harvest your virus. The system provides enough reagents for 16 packaging reactions.

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.

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.