Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) gives you the flexibility to easily produce high-titer lentivirus pseudotyped with any SARS-CoV-2 spike variant desired. Simply clone any desired SARS-CoV-2 spike variant sequence into the provided expression vector, dilute in sterile water with your transfer vector of choice, and add to a tube of single shots. Then, apply the reconstituted transfection mix to packaging cells in a 10-cm dish.
For your convenience, three plasmids encoding the truncated SARS-CoV-2 spike variants D614G, B.1.1.7, and B.1.351 are included. Additional SARS-CoV-2 spike variant mutants can be created using the provided spike variant sequences as template along with the supplied pEmpty vector in a simple, one-step In-Fusion Snap Assembly reaction.
Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots (Universal).
Transduction of an ACE2-positive cell line with SARS-CoV-2 pseudovirus. Lenti-X Packaging Single Shots (Universal) was used to produce truncated SARS-CoV-2 spike B.1.1.7 pseudovirus encoding the fluorescent protein ZsGreen (Panel A) or SARS-CoV-2 spike B.1.1.7 pseudovirus encoding firefly luciferase (Panel B). Panel A. Concentrated supernatant (20X; 30 or 10 µl) was used to transduce a HEK293T cell line stably expressing the human ACE2 receptor in the presence of 6 µg/ml polybrene in 48-well plates. The transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction. The percentage of ZsGreen-positive (ZsGreen+) cells and mean fluorescence intensity (MFI) are shown. Panel B. Concentrated supernatant (20X; 30, 20, or 10 µl) was used to transduce a HEK293T cell line stably expressing the human ACE2 receptor in the presence of 6 µg/ml polybrene in 48-well plates. HEK293T cells lacking the ACE2 transgene were transduced to determine background luminescence levels (NC; negative control). Luminescence values for each sample were measured 6 days post-transduction.
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Skip the PCR and directly use double-stranded DNA (dsDNA) fragments to seamlessly clone any desired SARS-CoV-2 spike variant with In-Fusion Snap Assembly. Our tutorial walks you through how to design In-Fusion-ready dsDNA fragments using the vectors provided with the Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) kit.
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Tutorial: Cloning dsDNA fragments as an alternative to PCR products to create SARS-CoV-2 spike variant mutants
Skip the PCR and directly use double-stranded DNA (dsDNA) fragments to seamlessly clone any desired SARS-CoV-2 spike variant with In-Fusion Snap Assembly. Our tutorial walks you through how to design In-Fusion-ready dsDNA fragments using the vectors provided with the Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) kit.
