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Product Overview

Producing high titers of replication-incompetent retrovirus

Transfect—Collect—Infect

  • Obtain retrovirus in 48 hours
  • Consistently high titers
  • Choice of envelope protein

The Retro-X Universal Packaging System provides a rapid and convenient method for producing high titers of replication-incompetent retrovirus. Choose from four different envelope proteins to package pseudotyped retrovirus with broad tropism to infect most commonly used cell types.

Introduction Results Conclusions

Introduction  

How the Retro-X Universal Packaging System works

Retroviral gene transfer systems depend on packaging cell lines to supply necessary viral structural proteins. The provided GP2-293 cell line contains the Moloney Murine Leukemia Virus (MMLV) gag and pol genes stably integrated into its genome. Simply transfect GP2-293 cells with the retroviral vector and the appropriate viral envelope vector, wait 48 hours, and then collect your viral supernatant which is ready for transduction (Figure 1).

The Retro-X Universal Packaging System

Figure 1. The Retro-X Universal Packaging System. The system consists of the GP2-293 Packaging Cell Line, which expresses viral Gag and Pol proteins, and a set of 4 viral envelope expression vectors. Cotransfecting an envelope vector with your retroviral construct allows you to select the ideal tropism for your virus.

Results  

Produce high titer retrovirus for your cell type

Broad tropism—infect all your cells

Viral envelopes are classified by the receptors they use to enter host cells. For example, ecotropic viruses are recognized by receptors only found on mouse and rat cells. The universal packaging system includes four envelope expression vectors—ecotropic, amphotropic, dualtropic (10A1), and pantropic (VSV-G)—allowing you to select the most appropriate envelope protein to infect your cell type (Table 1).

Target cell infectivity of Retrovirus

Obtain high titers of retroviral particles

The included cell line, GP2-293, has been optimized for the production of high-titer retrovirus packaged in any of the four envelope proteins (Figure 2). Rapid and easy determination of retroviral titer may be achieved using the Retro-X qRT-PCR Titration Kit (sold separately). If desired, retroviral supernatants are easily concentrated with the Retro-X Concentrator (not shown).

High-titer retroviral packaging in GP2-293 cells

Figure 2. High-titer retroviral packaging in GP2-293 cells. Cells were transfected with a retroviral vector expressing the red fluorescent protein DsRed2, and each of the four envelope vectors supplied with the Retro-X Universal Packaging System (Panels A–D). After incubating the transfected GP2-293 cells for 48 hr, 10 μl of the resulting virus was used to infect NIH 3T3 cells. After 48 hours, titers were measured by flow cytometry. Panel A. Ecotropic envelope. Panel B. Amphotropic envelope. Panel C. 10A1 envelope. Panel D. VSV-G envelope. Positive cells are indicated by the blue-filled line. Negative control cells are indicated by the purple line.

Conclusions  

The Retro-X Universal Packaging System provides a rapid and convenient method for producing high titers of replication-incompetent retrovirus. It includes four envelope expression vectors—ecotropic, amphotropic, dualtropic (10A1), and pantropic (VSV-G)—allowing you to select the most appropriate envelope protein to infect your cell type.

Related products

Cat. # Product Size Price License Quantity Details
631530 Retro-X™ Universal Packaging System Each $820.00

License Statement

ID Number  
8 For Research Use Only, Not for Use in Humans.

This kit provides a complete retroviral vector packaging system for use with any retroviral vector. It is designed to produce virus that can infect dividing cells from a broad range of mammalian and nonmammalian cell types. The GP2-293 Packaging Cell Line contains only the MoMuLV gag and pol genes. The viral envelope portion of the packaging function (env gene) is supplied by transiently co-transfecting one of the Retro-X Universal Packaging Vectors (p10A1, pAmpho, pEco, or pVSV-G) with the retroviral vector. The pQCLIN Retroviral Vector (included in the vector set) can be used as a positive control to constitutively express β-galactosidase in transduced mammalian cells.

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 You May Also Like Image Data

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Schematic for retroviral packaging using Retro-X Universal Packaging System

Schematic for retroviral packaging using Retro-X Universal Packaging System
Schematic for retroviral packaging using Retro-X Universal Packaging System.

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Retrovirus packaging options from Clontech

Retrovirus packaging options from Clontech
Retrovirus packaging options from Clontech. Summary of the cell lines and tropisms associated with commonly used retroviral envelopes.

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Characteristics of retroviral packaging cell lines: AmphoPack-293, EcoPack 2-293, and RetroPack PT67

Characteristics of retroviral packaging cell lines: AmphoPack-293, EcoPack 2-293, and RetroPack PT67
Characteristics of retroviral packaging cell lines: AmphoPack-293, EcoPack 2-293, and RetroPack PT67.

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High titer retroviral packaging with a choice of four different retroviral packaging envelopes

High titer retroviral packaging with a choice of four different retroviral packaging envelopes

High titer retroviral packaging with a choice of four different retroviral packaging envelopes. GP2-293 Cells were transfected with a Retro-X Q vector expressing the red fluorescent protein DsRed2, and each of the four envelope vectors supplied with the Retro-X Unversal Packaging System. After incubating the transfected GP2-293 cells for 48 hr, 10 ?l of the resulting virus was used to infect NIH 3T3 cells. After 48 hours, titers were measured by flow cytometry. Titers were calculated by multiplying the percent infected cells (infectious units) by the total cell number, then dividing by the volume of virus (ml). Panel A. Ecotropic envelope. Panel B. Amphotropic envelope. Panel C. Dualtropic envelope. Panel D. VSV-G envelope (pantropic). Positive cells are indicated by the red line. Negative control cells are indicated by the green line.


Learn more about our retroviral systems

Retroviral product selection guide

A selection guide to help you choose the best retroviral vectors, packaging systems, and other tools.

Retro-X FAQs

Find answers to your questions related to recombinant retrovirus experiments.

Universal packaging system

Versatile packaging system for producing high titers of virus with a choice of commonly used envelopes.

Retro-X Concentrator

Increase your viral titer up to 100-fold with Retro-X Concentrator—without ultracentrifugation.

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Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, 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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Capturem Trypsin for a rapid, efficient mass spectometry workflow at room temperature.

Speed up your mass spec workflow

Capturem Trypsin provides rapid, efficient, and complete digestion of protein samples, allowing an uninterrupted mass spectometry workflow at room temperature for downstream protein analysis. This product utilizes our novel Capturem technology in a spin column format with membrane-immobilized trypsin. Capturem Trypsin Columns may be used to completely digest protein samples in less than a minute with digestion efficiencies (protein coverage) comparable to or better than those obtained using in-solution trypsin digestion.

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