SARS-CoV-2 pseudovirus packaging systems - B.1.351
Lenti-X SARS-CoV-2 Packaging Single Shots enable streamlined, high-titer production of lentiviral particles pseudotyped with SARS-CoV-2 spike protein; simply add water containing your transfer vector of choice to a tube of single shots and then apply the reconstituted transfection mix to packaging cells in a 10-cm dish. For your convenience, Lenti-X SARS-CoV-2 packaging mixes are sold separately or bundled with ZsGreen1, luciferase transfer vectors, and negative control packaging reagents.
Lenti-X SARS-CoV-2 Packaging Single Shots enable streamlined, high-titer production of lentiviral particles pseudotyped with SARS-CoV-2 spike protein; simply add water containing your transfer vector of choice to a tube of single shots and then apply the reconstituted transfection mix to packaging cells in a 10-cm dish. For your convenience, Lenti-X SARS-CoV-2 packaging mixes are sold separately or bundled with ZsGreen1, luciferase transfer vectors, and negative control packaging reagents.
Separate Lenti-X packaging mixes are available for production of pseudovirus bearing full-length or truncated forms of the spike protein from the SARS-CoV-2 variant B.1.351 (originally identified in South Africa). The truncation involves removal of 19 amino acids encoding an ER retention signal on the protein C-terminus, and is associated with increased abundance of spike protein on the resulting particles, and increased particle infectivity (Johnson et al. 2020).
Overview
- High-performance packaging system consistently yields high titers: avoid spending time optimizing reagents, focus on your downstream assays
- Convenient single shots format minimizes hands-on time and likelihood of errors: simply add water containing a transfer plasmid of choice and apply to packaging cells
- Packaging mixes sold separately or as bundles: conserve resources, purchase only what you need
- Controls provided for neutralization assays: use the included packaging mix for production of lentiviral particles lacking an envelope protein
- Additional packaging mixes available for different SARS-CoV-2 spike protein variants: D614G, Wuhan-Hu-1
- Accompanying ACE2 cell line also available
More Information
Streamlined, one-step workflow
Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.
Consistently high titers
Transduction efficiencies and infectious titers using Lenti-X SARS-CoV-2 Packaging Single Shots. Lenti-X SARS-CoV-2 Packaging Single Shots (B.1.351 or D614G spike, full length or truncated) were used to produce pseudovirus encoding the fluorescent protein ZsGreen1. Viral particles were concentrated 50-fold (B.1.351 full length) or 20-fold (B.1.351 truncated and D614G) using Lenti-X Concentrator following the standard protocol, and different volumes of each prep were 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction (Panel A) and functional titers were calculated (Panel B).
Efficient transduction of ACE2-positive cells
Transduction of ACE2 HEK293T cells using B.1.351 SARS-CoV-2 pseudovirus encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots (B.1.351 spike, full length or truncated) were used to produce pseudovirus encoding firefly luciferase. Viral particles were concentrated 20-fold using Lenti-X Concentrator following the standard protocol, and different volumes of each prep were used to transduce an HEK293T cell line stably expressing the human ACE2 receptor in the presence of 6 µg/ml polybrene in 48-well plates. HEK293T cells lacking ACE2 expression were transduced in parallel to determine background luminescence levels, since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2. Luminescence values for each sample were measured 3 days posttransduction.
Ideal for neutralization studies
Neutralizing activity of soluble ACE2 protein against pseudoviruses bearing SARS-CoV-2 spike protein variants. Panels A and B. Serial dilutions of soluble ACE2 protein fused to the Fc domain from IgG (ACE2-Fc) were applied, along with SARS-CoV-2 pseudovirus, to ACE2 HEK293T cells. Luciferase levels were measured 3 days post-infection as a readout for virus infectivity. Data are graphed as percent neutralization relative to virus-only control infection. Values are mean ±SD and experiments were performed in triplicate.
References
Johnson, MC. et al. Optimized Pseudotyping Conditions for the SARS-COV-2 Spike Glycoprotein. J Virol. 94, e01062-20 (2020).
Additional product information
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.
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