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Simplified production of SARS-CoV-2-pseudotyped lentivirus

Lenti-X

A common objective in developing COVID-19 therapies is to demonstrate inhibition of SARS-CoV-2 infection. Given the safety measures required in handling live SARS-CoV-2, researchers often employ alternative viruses pseudotyped with the SARS-CoV-2 spike protein (Ni et al. 2020, Shang et al. 2020). To support these efforts, we have adapted our Lenti-X packaging system to enable high-titer production of spike-pseudotyped lentiviral particles in a convenient format.

A common objective in developing COVID-19 therapies is to demonstrate inhibition of SARS-CoV-2 infection. Given the safety measures required in handling live SARS-CoV-2, researchers often employ alternative viruses pseudotyped with the SARS-CoV-2 spike protein (Ni et al. 2020, Shang et al. 2020). To support these efforts, we have adapted our Lenti-X packaging system to enable high-titer production of spike-pseudotyped lentiviral particles in a convenient format.

Lenti-X SARS-CoV-2 Packaging Single Shots kits include lyophilized mixtures of Xfect transfection reagent and lentiviral packaging plasmids bearing any of four different spike protein variants, accompanying transfer vectors encoding ZsGreen1 and luciferase reporters, and transfection mixes for production of lentiviral particles lacking an envelope protein (commonly employed as negative controls). High titers of SARS-CoV-2 pseudovirus are obtained by a simple, one-step method: just add water containing your transfer vector of choice to a tube of single shots, and then apply the reconstituted transfection mix to packaging cells (e.g., Lenti-X 293T cells) in a 10-cm dish. Viral supernatants can be harvested 48–72 hours posttransfection.

Packaging systems are available for the production of viral particles bearing your choice of four different SARS-CoV-2 spike protein variants:

  • WT (full length)—derived from the Wuhan-Hu-1 isolate (NC_045512.2) of the novel coronavirus SARS-CoV-2, codon-optimized for expression in mammalian cells
  • D614G (full length)—includes the D614G mutation in the spike protein coding sequence; the prevalence of this mutation has risen rapidly since the start of the COVID-19 pandemic (Isabel et al. 2020)
  • WT (truncated) and D614G (truncated)—truncations of WT spike and D614G spike coding sequences involving removal of 19 amino acids from C-termini; the truncation is associated with increased abundance of spike protein on the resulting particles, and increased particle infectivity (Johnson et al. 2020)

For your convenience, Lenti-X SARS-CoV-2 packaging mixes are also sold separately.

 More  Less
Cat. # Product Size Price License Quantity Details
632673 Lenti-X™ SARS-CoV-2 Packaging Mix (D614G Spike, Full Length) 12 Rxns USD $795.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length) provides an extremely simple and consistent one-step method for producing high-titer lentivirus pseudotyped with the spike protein from the SARS-CoV-2 virus (D614G mutant). No additional transfection reagent is needed because Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length) consists of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral transfer vector of choice in sterile water and adding it to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Cat. # 632673 includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length) and is also sold as part of Lenti-X SARS-CoV-2 Packaging Single Shots (D614G Spike, Full Length; Cat. # 632669).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

632673: Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length)

632673: Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length)

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632675 Lenti-X™ SARS-CoV-2 Packaging Mix (D614G Spike, Truncated) 12 Rxns USD $795.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated) provides an extremely simple and consistent one-step method for producing high-titer lentivirus pseudotyped with a truncated form of the spike protein from the SARS-CoV-2 virus (D614G mutant). No additional transfection reagent is needed because Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated) consists of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral transfer vector of choice in sterile water and adding it to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Cat. # 632675 includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated) and is also sold as part of Lenti-X SARS-CoV-2 Packaging Single Shots (D614G Spike, Truncated; Cat. # 632671).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

632675: Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated)

632675: Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated)

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (truncated form) and encoding the fluorescent protein ZsGreen1. 100 μl of supernatant from each prep was used to transduce the Human ACE2 293T Cell Line. Panel A. Microscopy images of transduced cells at 72 hours post-infection. Panel B. Transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632676 Lenti-X™ SARS-CoV-2 Packaging Mix (No-Envelope Control) 6 Rxns USD $395.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control) provides an extremely simple and consistent one-step method for producing high-titer lentivirus lacking an envelope protein. No additional transfection reagent is needed because Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control) consists of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral transfer vector of choice in sterile water and adding it to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Cat. # 632676 includes 6 tubes of Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control) and is also sold as part of Lenti-X SARS-CoV-2 Packaging Single Shots (Cat. #s 632668, 632669, 632670, 632671).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

632676: Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control)

632676: Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control)

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632672 Lenti-X™ SARS-CoV-2 Packaging Mix (WT Spike, Full Length) 12 Rxns USD $795.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length) provides an extremely simple and consistent one-step method for producing high-titer lentivirus pseudotyped with the spike protein from the SARS-CoV-2 virus (Wuhan-Hu-1 isolate). No additional transfection reagent is needed because Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length) consists of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral transfer vector of choice in sterile water and adding it to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Cat. # 632672 includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length) and is also sold as part of Lenti-X SARS-CoV-2 Packaging Single Shots (WT Spike, Full Length; Cat. # 632668).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

632672: Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length)

632672: Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length)

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632674 Lenti-X™ SARS-CoV-2 Packaging Mix (WT Spike, Truncated) 12 Rxns USD $795.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated) provides an extremely simple and consistent one-step method for producing high-titer lentivirus pseudotyped with a truncated form of the spike protein from the SARS-CoV-2 virus (Wuhan-Hu-1 isolate). No additional transfection reagent is needed because Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated) consists of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral transfer vector of choice in sterile water and adding it to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Cat. # 632674 includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated) and is also sold as part of Lenti-X SARS-CoV-2 Packaging Single Shots (WT Spike, Truncated; Cat. # 632670).

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

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Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

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Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

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632674: Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated)

632674: Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated)

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Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (truncated form) and encoding the fluorescent protein ZsGreen1. 100 μl of supernatant from each prep was used to transduce the Human ACE2 293T Cell Line. Panel A. Microscopy images of transduced cells at 72 hours post-infection. Panel B. Transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction.

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Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

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Neutralization data with convalescent serum

Neutralization data with convalescent serum

Use of the Human ACE2 293T Cell Line in neutralization assays employing pooled non-human primate convalescent serum to SARS-CoV-2 as a blocking agent. WT (truncated) SARS-CoV-2 pseudovirus was incubated with serial dilutions of the convalescent serum (BEI Resources, Cat. # NR-52401) and applied to the Human ACE2 293T Cell Line. 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. Negative control was performed using serum from healthy macaques

632669 Lenti-X™ SARS-CoV-2 Packaging Single Shots (D614G Spike, Full Length) 12 Rxns USD $975.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Single Shots (D614G Spike, Full Length) enable streamlined production of high-titer lentivirus pseudotyped with the spike protein from SARS-CoV-2 (D614G mutant) by providing pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of Lenti-X lentiviral packaging plasmids. Pseudovirus production simply involves reconstituting a tube of lyophilized packaging mix with a lentiviral transfer vector of choice in sterile water and adding the mixture to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Each kit includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Full Length; Cat. # 632673), 6 tubes of Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control; Cat. # 632676) for production of pseudoviral particles lacking an envelope protein, and the Lenti-X SARS-CoV-2 Vector Set (Cat. # 632677), which includes two self-inactivating lentiviral transfer vectors encoding ZsGreen1 and firefly luciferase, respectively, under the CMV promoter, to be used as reporters for viral transduction.

For your convenience, the packaging mixes included in the kit are also sold separately.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

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Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

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Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

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Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632671 Lenti-X™ SARS-CoV-2 Packaging Single Shots (D614G Spike, Truncated) 12 Rxns USD $975.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Single Shots (D614G Spike, Truncated) enable streamlined production of high-titer lentivirus pseudotyped with a truncated form of the spike protein from SARS-CoV-2 (D614G mutant) by providing pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of Lenti-X lentiviral packaging plasmids. Pseudovirus production simply involves reconstituting a tube of lyophilized packaging mix with a lentiviral transfer vector of choice in sterile water and adding the mixture to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Each kit includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (D614G Spike, Truncated; Cat. # 632675), 6 tubes of Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control; Cat. # 632676) for production of pseudoviral particles lacking an envelope protein, and the Lenti-X SARS-CoV-2 Vector Set (Cat. # 632677), which includes two self-inactivating lentiviral transfer vectors encoding ZsGreen1 and firefly luciferase, respectively, under the CMV promoter, to be used as reporters for viral transduction.

For your convenience, the packaging mixes included in the kit are also sold separately.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

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Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

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Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (truncated form) and encoding the fluorescent protein ZsGreen1. 100 μl of supernatant from each prep was used to transduce the Human ACE2 293T Cell Line. Panel A. Microscopy images of transduced cells at 72 hours post-infection. Panel B. Transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction.

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Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632668 Lenti-X™ SARS-CoV-2 Packaging Single Shots (WT Spike, Full Length) 12 Rxns USD $975.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Single Shots (WT Spike, Full Length) enable streamlined production of high-titer lentivirus pseudotyped with the spike protein from SARS-CoV-2 (Wuhan-Hu-1 isolate) by providing pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of Lenti-X lentiviral packaging plasmids. Pseudovirus production simply involves reconstituting a tube of lyophilized packaging mix with a lentiviral transfer vector of choice in sterile water and adding the mixture to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Each kit includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Full Length; Cat. # 632672), 6 tubes of Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control; Cat. # 632676) for production of pseudoviral particles lacking an envelope protein, and the Lenti-X SARS-CoV-2 Vector Set (Cat. # 632677), which includes two self-inactivating lentiviral transfer vectors encoding ZsGreen1 and firefly luciferase, respectively, under the CMV promoter, to be used as reporters for viral transduction.

For your convenience, the packaging mixes included in the kit are also sold separately.

 

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

632670 Lenti-X™ SARS-CoV-2 Packaging Single Shots (WT Spike, Truncated) 12 Rxns USD $975.00

License Statement

ID Number  
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 6,998,115; 7,427,394. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This product is protected by Japanese Patent No. 6454352 and corresponding U.S. pending patent and other foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at  licensing@takarabio.com.

Lenti-X SARS-CoV-2 Packaging Single Shots (WT Spike, Truncated) enable streamlined production of high-titer lentivirus pseudotyped with a truncated form of the spike protein from SARS-CoV-2 (Wuhan-Hu-1 isolate) by providing pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of Lenti-X lentiviral packaging plasmids. Pseudovirus production simply involves reconstituting a tube of lyophilized packaging mix with a lentiviral transfer vector of choice in sterile water and adding the mixture to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Each kit includes 12 tubes of Lenti-X SARS-CoV-2 Packaging Mix (WT Spike, Truncated; Cat. # 632674), 6 tubes of Lenti-X SARS-CoV-2 Packaging Mix (No-Envelope Control; Cat. # 632676) for production of pseudoviral particles lacking an envelope protein, and the Lenti-X SARS-CoV-2 Vector Set (Cat. # 632677), which includes two self-inactivating lentiviral transfer vectors encoding ZsGreen1 and firefly luciferase, respectively, under the CMV promoter, to be used as reporters for viral transduction.

For your convenience, the packaging mixes included in the kit are also sold separately.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.

Back

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein from SARS-CoV-2, respectively, and encoding the fluorescent protein ZsGreen1. Experiments were performed in duplicate. 100 μl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Following the same protocol, pseudoviruses encoding firefly luciferase were produced in duplicates and used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity since spike-pseudotyped viruses are incapable of infecting HEK293T cells in the absence of ACE2). Resulting levels of luciferase were measured 6 days posttransduction.

Back

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with pseudovirus produced using Lenti-X SARS-CoV-2 Packaging Single Shots (WT or D614G spike, Truncated). Panel B. Corresponding microscopy images of ACE2 HEK293T cells transduced with SARS-CoV-2 pseudoviruses encoding the ZsGreen1 reporter at 72 hours post-infection.

Back

Neutralization data with ACE2 protein as a blocking agent

Neutralization data with ACE2 protein as a blocking agent

Use of the Human ACE2 293T Cell Line in neutralization assays employing soluble ACE2 protein as a blocking agent. 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 the Human ACE2 293T Cell Line. 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.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding ZsGreen1. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (truncated form) and encoding the fluorescent protein ZsGreen1. 100 μl of supernatant from each prep was used to transduce the Human ACE2 293T Cell Line. Panel A. Microscopy images of transduced cells at 72 hours post-infection. Panel B. Transduction efficiencies for each sample were measured by flow cytometry 6 days post-transduction.

Back

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase

Transduction of Human ACE2 293T Cell Line with SARS-CoV-2 pseudoviruses encoding luciferase. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT or D614G variants of the spike protein (full length or truncated forms) and encoding the firefly luciferase protein. 10 μl of concentrated virus (21X) from each prep was used to transduce the Human ACE2 293T Cell Line. Luciferase activity was measured 6 days post-transduction using a control virus lacking an envelope protein to provide a background signal for analysis of the luciferase activity.

Back

Neutralization data with convalescent serum

Neutralization data with convalescent serum

Use of the Human ACE2 293T Cell Line in neutralization assays employing pooled non-human primate convalescent serum to SARS-CoV-2 as a blocking agent. WT (truncated) SARS-CoV-2 pseudovirus was incubated with serial dilutions of the convalescent serum (BEI Resources, Cat. # NR-52401) and applied to the Human ACE2 293T Cell Line. 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. Negative control was performed using serum from healthy macaques

Human ACE2 stable cell line
Check out Lenti-X GoStix for instant titer measurement of lentiviral titer Lenti-X GoStix Plus product page
Lenti-X Concentrator

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
  • Packaging mixes available for different SARS-CoV-2 spike protein variants: choose from the following four options:
    • WT (full length)—derived from the Wuhan-Hu-1 isolate (NC_045512.2)
    • D614G (full length)—coding sequence includes the highly prevalent D614G mutation
    • WT (truncated) and D614G (truncated)—truncated coding sequences result in deletion of 19 amino acids from spike C-terminus and are associated with increased particle infectivity
  • Controls provided for neutralization assays: use the included packaging mix for production of lentiviral particles lacking an envelope protein

More Information

Streamlined, one-step workflow

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots

Workflow for pseudovirus production using Lenti-X SARS-CoV-2 Packaging Single Shots.


Efficient transduction of ACE2-positive cells

Transduction of an ACE2-positive cell line using pseudovirus produced with Lenti-X SARS-CoV-2 Packaging Single Shots

Transduction of an ACE2-positive cell line with SARS-CoV-2 pseudovirus. Panel A. Lenti-X SARS-CoV-2 Packaging Single Shots were used to generate lentiviral particles pseudotyped with either WT (Wuhan-Hu-1) or D614G variants of the spike protein, and encoding the fluorescent protein ZsGreen1. 100 µl of supernatant from each prep was 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. Transduction efficiencies for each sample were measured by flow cytometry 6 days posttransduction. Panel B. Pseudoviruses encoding firefly luciferase were used to transduce both the ACE2-positive cell line and an HEK293T cell line lacking ACE2 expression (included to provide a background signal for analysis of luciferase activity). Luciferase activity was measured 6 days posttransduction.


Consistently high titers

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots

Functional titers obtained using Lenti-X SARS-CoV-2 Packaging Single Shots. Panel A. Functional titers determined via analysis of ZsGreen1 reporter expression in ACE2-positive HEK293T cells transduced with SARS-CoV-2 pseudovirus. Panel B. Corresponding microscopy images of transduced ACE2-positive cells at 72 hours post-infection.


Ideal for neutralization studies

Neutralizing activity of soluble ACE2 protein fused to the Fc domain from IgG (ACE2-Fc) against SARS-CoV-2 pseudoviruses bearing truncated WT or D614G spike protein variants

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

Ni, L. et al. Detection of SARS-CoV-2-Specific Humoral and Cellular Immunity in COVID-19 Convalescent Individuals. Immunity. 52, 971–977 (2020).

Shang, J. et al. Cell entry mechanisms of SARS-CoV-2. Proc Natl Acad Sci USA. 117, 11727–11734 (2020).

Isabel, S. et al. Evolutionary and structural analyses of SARS-CoV-2 D614G spike protein mutation now documented worldwide. Sci Rep. 10, 14031 (2020).

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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