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  • Universal
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Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) tech note
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Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) tech note

SARS-CoV-2 pseudovirus packaging systems - universal

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.

Cat. # Product Size License Quantity Details
631295 Lenti-X™ SARS-CoV-2 Packaging Single Shots (Universal) 16 Rxns USD $1095.00

License Statement

ID Number  
55 cPPT Element. This product and its use are the subject to one or more of the following U.S. Pat. No. 8,093,042. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA, Inc. is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).
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 (Universal) allow streamlined production of high-titer lentivirus pseudotyped with your SARS-CoV-2 spike protein of interest 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 involved diluting a SARS-CoV-2 spike variant expression vector and your transfer vector of choice in water, adding to a tube of single shots, and applying the mixture to 293T cells (e.g., Lenti-X 293T Cells, Cat. # 632180) in a 10-cm dish.

Each kit includes 16 tubes of Lenti-X Packaging Single Shots (Envelope-Free; Cat. # 631297), the pEmpty Vector (Cat. # 631298) for cloning and expression of the desired SARS-CoV-2 spike protein, the Lenti-X SARS-CoV-2 Spike Vector Set (Cat. # 631296), which includes three plasmids encoding for truncated SARS-CoV-2 spike variants D614G, B.1.1.7, and B.1.351 and the Lenti-X Reporter Vector Set (Cat. # 632677). Cat. # 632677 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.

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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.

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

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

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

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

Transduction of an ACE2-positive cell line using pseudovirus produced with 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.

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Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) uses Takara Bio’s Lenti-X 4th-generation packaging system

Lenti-X SARS-CoV-2 Packaging Single Shots (Universal) uses Takara Bio’s Lenti-X 4th-generation packaging system

Lenti-X Packaging Single Shots (Envelope-Free) uses Takara Bio’s Lenti-X 4th-generation packaging system. Panel A. Plasmids expressing all viral components except the SARS-CoV-2 spike protein that are contained in lyophilized Single Shots formulation. High expression of essential viral components are driven by the Tet-Off and Tat transactivators. In addition, the gag and pol genes are separated and the pol gene is fused to vpr to ensure transport of the reverse transcriptase/integrase protein into the recombinant viral particle. Panel B. The remaining viral component, the desired SARS-CoV-2 spike variant mutant is cloned into the pEmpty Vector and used in the single shots reaction. Expression of the spike variant mutant is driven by a CMV promoter.

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631295: Lenti-X SARS-CoV-2 Packaging Single Shots (Universal)

631295: Lenti-X SARS-CoV-2 Packaging Single Shots (Universal)

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

  • Customizable SARS-CoV-2 pseudovirus: produce any SARS-CoV-2 spike protein variant you need for your research
  • Simple cloning procedure: the provided pEmpty Vector paired with In-Fusion Snap Assembly makes spike variant protein plasmid production a breeze
  • 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 the SARS-CoV-2 spike variant vector and lentiviral transfer vector, and apply to packaging cells
  • B.1.351, B.1.1.7, and D614G SARS-CoV-2 spike protein variant plasmids included: use directly in the Lenti-X SARS-CoV-2 Single Shots (Universal) reaction or as template for the creation of additional SARS-CoV-2 spike variant mutants
  • 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 (Universal).


Efficient transduction of ACE2-positive cells

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.


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.


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.

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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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.

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