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AAV2 tetracycline-inducible expression system

AAV2 Tetracycline Inducible Expression System

Tet-One systems provide all necessary components for tetracycline-inducible (tet) expression on a single vector delivered by AAV2 virus. The Tet-On 3G transactivator is expressed from the human phosphoglycerate kinase 1 (PGK) promoter, and the cloned gene of interest is expressed from the PTRE3GS promoter. The AAVpro system contains the reagents to generate the Tet-One vector, as well as components to efficiently package and isolate AAV2 particles.

Tet-One systems provide all necessary components for tetracycline-inducible (tet) expression on a single vector delivered by AAV2 virus. The Tet-On 3G transactivator is expressed from the human phosphoglycerate kinase 1 (PGK) promoter, and the cloned gene of interest is expressed from the PTRE3GS promoter. The AAVpro system contains the reagents to generate the Tet-One vector, as well as components to efficiently package and isolate AAV2 particles. This kit uses helper-free packaging, a unique system for preparing high-titer AAV2 particles without the use of a helper virus.

There are also several other Tet-One vector options for mammalian cells.

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Cat. # Product Size License Quantity Details
634310 AAVpro® Tet-One™ Inducible Expression System (AAV2) 1 System USD $976.00

License Statement

ID Number  
42 Use of the Tetracycline controllable expression systems (the "Tet Technology") is covered by a series of patents including U.S. Patent # 8383364, # 9181556 , European patents EP # 1954811, #2352833 and corresponding patent claims outside these regions which are proprietary to TET Systems GmbH & Co. KG. Academic research institutions are granted an automatic license with the purchase of this product to use the Tet Technology only for internal, academic research purposes, which license specifically excludes the right to sell, or otherwise transfer, the Tet Technology or its component parts to third parties. Notwithstanding the above, academic and not-for profit research institutions whose research using the Tet Technology is sponsored by for profit organizations, which shall receive ownership to any data and results stemming from the sponsored research, shall need a commercial license agreement from TET Systems in order to use the Tet Technology. In accepting this license, all users acknowledge that the Tet Technology is experimental in nature. TET Systems GmbH & Co. KG makes no warranties, express or implied or of any kind, and hereby disclaims any warranties, representations, or guarantees of any kind as to the Tet Technology, patents, or products. All others are invited to request a license from TET Systems GmbH & Co. KG prior to purchasing these reagents or using them for any purpose. Takara Bio USA, Inc. is required by its licensing agreement to submit a report of all purchasers of the Tet-controllable expression system to TET Systems.

For license information, please contact:
GSF/CEO
TET Systems GmbH & Co. KG,
Im Neuenheimer Feld 582
69120 Heidelberg
Germany
Tel: +49 6221 5880400
Fax: +49 6221 5880404
email: info@tetsystems.com
or use the electronic licensing request form via http://www.tetsystems.com/ip-licensing/licensing/for-profit-research
M83 This product is the subject of the claims of US patent No. 9422576 and its foreign counterparts.
M84 This product is the subject of the claims of US patent No. 10072250 and its foreign counterparts.

The AAVpro Tet-One Inducible Expression System is a tetracycline-inducible gene expression system that allows you to produce high titers of recombinant, adeno-associated virus (AAV) for establishing a tightly inducible expression system for your gene of interest in mammalian cells. The all-in-one pAAV-TetOne vector expresses the Tet-On 3G transactivator from the constitutive human PGK promoter, and your gene of interest from the PTRE3GS promoter. In the presence of doxycycline (Dox), the Tet-On 3G transactivator specifically binds and activates high-level transcription from the inducible promoter that controls expression of your gene. There is no selection marker on this vector for mammalian cells. The kit also includes the plasmids for preparation of high-titer AAV particles without the use of a helper virus, and AAV Extraction Solution for isolation of pure AAV particles.

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.

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Establishing an inducible expression system in target cells with AAVpro Tet-One

Establishing an inducible expression system in target cells with AAVpro Tet-One
Establishing an inducible expression system in target cells with AAVpro Tet-One. The pAAV-TetOne Vector containing your gene of interest, pRC2-mi342, and pHelper are cotransfected into HEK 293 or HEK 293T cell lines to generate a high-titer AAV particles. The AAV particles are used to transduce your target cells, and Dox is added to express your gene of interest when desired.

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Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV

Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV
Transduction and induction in dividing cell lines with different MOIs of TetOne-Luc Control AAV. 1 x 105 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with different MOIs (genomic titer), and 100 ng/ml Dox was added. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.

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Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox

Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox
Induction of luciferase after transduction with TetOne-Luc Control AAV in the presence of different concentrations of Dox. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and Dox was added at different concentrations. After 48 hr, cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer.

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Different induction times of luciferase after transduction with TetOne-Luc Control AAV

Different induction times of luciferase after transduction with TetOne-Luc Control AAV
Different induction times of luciferase after transduction with TetOne-Luc Control AAV. 1 x 105 HEK 293 cells were seeded in 12-well plates 12 hr before transduction. Cells were transduced with 1 x 105 MOI (genomic titer), and 100 ng/ml Dox was added. Cells were harvested and analyzed with a standard firefly luciferase assay using a luminometer at various time points after induction.

Required Products

Cat. # Product Size Price License Quantity Details
631101 Tet System Approved FBS, US-Sourced 500 mL USD $902.00

Fetal Bovine Serum (FBS) that has been functionally tested for optimal use with all of our Tet Systems. This FBS does not contain trace levels of tetracycline (or derivatives) which have been observed to interfere with proper regulation of TRE-controlled gene expression. Use of this serum ensures the maximal range of induction possible with the Tet Systems. This FBS has been processed in the United States.

Documents Components Image Data

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Fetal bovine serum (FBS) approved for use with tetracycline-inducible expression systems

Fetal bovine serum (FBS) approved for use with tetracycline-inducible expression systems

Fetal bovine serum (FBS) approved for use with tetracycline inducible expression systems. Only Takara Bio USA functionally tests FBS to be sure that no contaminating tetracycline derivatives interfere with induced expression levels.

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Our fetal bovine serum is functionally tested to guarantee consistent results with tetracycline-inducible expression systems

Our fetal bovine serum is functionally tested to guarantee consistent results with tetracycline-inducible expression systems

Fetal bovine serum from Clontech is functionally tested to guarantee consistent results with tetracycline-inducible expression systems. Serum from other vendors that have not been functionally tested may contain tetracyclines that affect the maximum expression in Tet-Off systems, or generate background in Tet-On systems. Data here shows induction of luciferase expression from our CHO-AA8-Luc Tet-Off Control Cell Line using different sources of FBS.

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631101: Tet System Approved FBS, US-Sourced

631101: Tet System Approved FBS, US-Sourced
631106 Tet System Approved FBS 500 mL USD $883.00

Fetal Bovine Serum (FBS) that has been functionally tested for optimal use with all of our Tet Systems. This FBS does not contain trace levels of tetracycline (or its derivatives) which have been observed to interfere with proper regulation of TRE-controlled gene expression. Use of this serum ensures the maximal range of induction possible with the Tet Systems.

Documents Components Image Data

Back

Fetal bovine serum (FBS) approved for use with tetracycline-inducible expression systems

Fetal bovine serum (FBS) approved for use with tetracycline-inducible expression systems

Fetal bovine serum (FBS) approved for use with tetracycline inducible expression systems. Only Takara Bio USA functionally tests FBS to be sure that no contaminating tetracycline derivatives interfere with induced expression levels.

Back

Our fetal bovine serum is functionally tested to guarantee consistent results with tetracycline-inducible expression systems

Our fetal bovine serum is functionally tested to guarantee consistent results with tetracycline-inducible expression systems

Fetal bovine serum from Clontech is functionally tested to guarantee consistent results with tetracycline-inducible expression systems. Serum from other vendors that have not been functionally tested may contain tetracyclines that affect the maximum expression in Tet-Off systems, or generate background in Tet-On systems. Data here shows induction of luciferase expression from our CHO-AA8-Luc Tet-Off Control Cell Line using different sources of FBS.

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631106: Tet System Approved FBS

631106: Tet System Approved FBS

Overviews Tet-One overview
Tips and FAQs AAV FAQs
Tips and FAQs Learn about Tet systems

Overview

  • All-in-one vector design allows you to establish tet-inducible expression of your gene of interest with a single vector
  • Low background expression and high induction
  • Transactivator expression from the human PGK promoter allows inducible expression in a broad range of tissues
  • The unique AAV packaging system allows preparation of high-titer AAV2 without the use of a helper virus
  • Kit includes AAV Extraction Solution that increases viral particle recovery as compared to conventional freeze-thaw methods

More Information

Applications

  • Tightly controlled, tetracycline-inducible expression of your gene of interest in mammalian cells
References

Heinz, N. et al. Retroviral and transposon-based tet-regulated all-in-one vectors with reduced background expression and improved dynamic range. Hum. Gene Ther. 22, 166–176 (2011).

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.


Tet-One tetracycline-inducible expression system, technology overview

Tighly controlled expression system that is as easy to use as any constitutive system

Until now, the best inducible mammalian expression systems have all required a two-vector format. Tet-One inducible expression systems are the only commercially available single-vector systems that can deliver a >10,000-fold difference in expression levels between induced and uninduced states.

Tet-One technology overview All inducible systems

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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, Takara Bio USA 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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