Simplify Lentiviral Packaging with Lenti-X Packaging Single Shots
In a single step, you can be on the way to high titers of VSV-G psuedotyped lentivirus. Lenti-X Packaging Single Shots are pre-aliquoted, lyophilized, single tubes that contain Xfect Transfection Reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids. This format provides greater simplicity, more consistent results, and ultra high lentiviral titers.
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Each Lenti-X Packaging Single Shots tube contains lyophilized transfection reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids (4th generation). Simply add your lentiviral vector of choice in sterile water and add to 293T cells in a 10-cm dish.
Eliminating variability during reaction set up leads to consistent transfections that consistently yield high viral titers.
Consistent, high-efficiency transfections lead to high titers. A lentiviral vector encoding ZsGreen1 was packaged according to the Lenti-X Single Shots protocol in four independent experiments. Forty-eight hours after transfection, the cells were imaged by fluorescence microscopy (Panel A, top) and light microscopy (Panel A, bottom). After images were taken, the supernatant was harvested and used to infect HT1080 cells for titer determination (Panel B, IFU/ml).
High viral titers
Lenti-X Packaging Single Shots can produce high titer virus regardless of the lentiviral vector backbone used and regardless of the method used for titration.
High-titer virus is produced regardless of the lentiviral vector backbone. A CMV ZsGreen1 expression cassette was cloned into several lentiviral vector backbones. These vectors were then packaged into lentivirus using the Lenti-X Packaging Single Shots following the provided protocol. After 48 hr, titer was determine using several methods. To determine infectivity, the supernatant was harvested and used to infect HT1080 cells (Flow Cytometry). Harvested viral supernatants were also analyzed by RT-PCR to quantify viral genome copies (qRT-PCR, Lenti-X qRT-PCR Titration Kit), by ELISA to measure p24 (p24 ELISA, Lenti-X p24 Rapid Titer Kit), and by a rapid lentiviral detection method (Lenti-X GoStix).