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Tetracycline-inducible adenovirus system

Tetracycline inducible adenovirus system

Takara Bio's Clontech-brand Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) combines the tightest and most sensitive control of gene expression with the most advanced commercially available adenoviral vector system. With this system, tightly controlled inducible expression is as easy as constitutive expression, and cloning into an adenoviral vector is as straightforward as cloning into any plasmid.

Takara Bio's Clontech-brand Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) combines the tightest and most sensitive control of gene expression with the most advanced commercially available adenoviral vector system. With this system, the tightly controlled inducible expression is as easy as constitutive expression, and cloning into an adenoviral vector is as straightforward as cloning into any plasmid.

How does the Tet-On 3G inducible system work?

Target cells that express the Tet-On 3G transactivator protein and contain a gene of interest (GOI) under the control of a TRE3G promoter (PTRE3G) will express high levels of your GOI, but only when cultured in the presence of doxycycline (Dox), a tetracycline analog. When bound by Dox, the Tet-On 3G protein undergoes a conformational change that allows it to bind to tet operator (tetO) sequences located in PTRE3G (please expand the Details tab in the product table for more information). In contrast to TetR-based systems, Tet-On technologies activate rather than repress transcription, a critical difference which results in far lower basal expression, higher maximal expression, a more rapid response time, and ultimately, the first choice for conditional expression.

What makes this system so easy to use?

  1. All-in-one vector—The Tet-On 3G transactivator gene has been precloned into the E3 region of the adenoviral genome and is expressed constitutively from a CMV promoter. Clone your gene of interest using In-Fusion HD at the E1 region of the adenovirus between the tightly regulated PTRE3G promoter and an SV40 polyA signal. Because the two regions are widely separated, interference from the CMV promoter cannot affect basal expression from PTRE3G, and so a very low basal expression and high fold-inducibility are retained (see data images by expanding the Details tab in the product table).
  2. Easy cloning—Until now the main drawback of commercially supplied adenoviral vector systems has been the need to use complex cloning procedures to overcome the difficulties with cloning into large (~34 kb) plasmids. Our Adeno-X virologists thought "wouldn't it be great if you could clone directly into the adenoviral plasmid just like any plasmid?" They then harnessed the power of In-Fusion HD Cloning technology to make this happen.

Lowest-ever background, highest sensitivity

The combination of two optimized elements makes Tet-On 3G the highest performing inducible expression system.

  1. PTRE3G promoter—Mutations have reduced background expression from the inducible promoter to very low levels compared to previous generations of the Tet-On system.
  2. Tet-On 3G transactivator protein—Compared to early generations, mutations have significantly increased its sensitivity to the inducer doxycycline. When the two elements are combined, not only can you detect high expression of your protein after exposure to Dox, you can control the level of expression by titration of the Dox concentration, and you can generate very high fold induction—up to a 3,000-fold difference between the induced and uninduced states. The maximum expression level can be manipulated by increasing the amount of virus per cell (see data included under the Details tab).

Unlike the leading competitor, Adeno-X really is easy

Compared to the leading competitor system, which requires 8 days or more for a cloning procedure that involves cloning into a shuttle vector and transformation of two different E. coli strains, the Adeno-X system is easy and allows you to finish cloning with high efficiency in just 2–3 days. Adeno-X uses no shuttle vector so requires no subcloning, and a single high-performance E. coli strain (Stellar) is included with the kit.

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Cat. # Product Size Price License Quantity Details
631180 Adeno-X™ Adenoviral System 3 (Tet-On® 3G Inducible) 10 Rxns $1868.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.
42 Use of the Tetracycline controllable expression systems (the "Tet Technology") is covered by a series of patents including U.S. Patent # 7541446, # 8383364, # 9181556 , European patents EP # 1200607, # 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
25 This product is covered by U.S. Patent No. 6,303,362.

Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) provides the components necessary to generate a tightly controlled, highly sensitive, inducible expression system in an all-in-one adenoviral vector format. pAdenoX-Tet3G (included in the vector set) is a prelinearized, adenoviral vector that is ready for the insertion of your gene of interest with our In-Fusion HD PCR Cloning technology. Simply PCR-amplify your gene and combine it with pAdenoX-Tet3G in an In-Fusion HD Cloning reaction. In-Fusion HD Cloning is fast, simple, precise and efficient, making Adeno-X Adenoviral System 3 the most advanced commercially available adenoviral gene delivery tool.

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 Resources

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The Adeno-X Tet-On 3G systems generate very high fold induction, with up to 3,000-fold difference between induced and uninduced states

The Adeno-X Tet-On 3G systems generate very high fold induction, with up to 3,000-fold difference between induced and uninduced states

The Adeno-X Tet-On 3G systems generate very high fold-induction, with up to 3,000-fold difference between induced and uninduced states. Using equal amounts of high-titer supernatants, HeLa cells cultured at the indicated concentrations of Dox were infected with Adeno-X Tet-On 3G luciferase virus. Cultures were harvested and assayed for luciferase activity.

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Expression increases with higher MOIs

Expression increases with higher MOIs

Expression increases with higher MOIs. HeLa cells were infected with varying MOIs of pAdenoX Tet-On 3G adenovirus that expresses luciferase. After four hours, the media was replaced with fresh media +/- doxycycline (1 μg/ml). Cultures were harvested and assayed for luciferase activity. Maximal expression increases with increasing MOI, which also results in a slight increase in background expression.

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The Adeno-X Tet-On 3G systems are highly inducible

The Adeno-X Tet-On 3G systems are highly inducible

The Adeno-X Tet-On 3G systems are highly inducible. Using equal amounts of high-titer supernatants, HeLa cells cultured at the indicated concentrations of Dox were infected with Adeno-X Tet-On 3G lacZ virus. Cultures were harvested and assayed for beta-galactosidase activity using the Luminescent Beta-galactosidase Reporter System 3 (Cat. # 631713).

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Adeno-X adenoviral system 3 (Tet-On 3G Inducible)

Adeno-X adenoviral system 3 (Tet-On 3G Inducible)

Adeno-X adenoviral system 3 (Tet-On 3G Inducible). Create a point mutation and clone in one step using In-Fusion Cloning.

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The Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) allows inducible gene expression only in the presence of doxycycline

The Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) allows inducible gene expression only in the presence of doxycycline

The Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) allows inducible gene expression only in the presence of doxycycline. The system includes In-Fusion HD Cloning Kit for cloning your gene of interest (GOI) directly into the easy-to-use, all-in-one pAdenoX-Tet3G expression vector.

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Comparison of Adeno-X adenoviral system 3 to the leading "Easy" competitor

Comparison of Adeno-X adenoviral system 3 to the leading "Easy" competitor

Comparison of Adeno-X adenoviral system 3 to the leading "Easy" competitor.

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Constructing recombinant adenovirus with In-Fusion Cloning technology

Constructing recombinant adenovirus with In-Fusion Cloning technology

Constructing recombinant adenovirus with In-Fusion Cloning technology. DNA sequences can be rapidly transferred as PCR products to any pAdenoX vector using the In-Fusion cloning method. In this example, your gene of interest is amplified with 15 bp extensions that are homologous to the ends of the linearized adenoviral vector. The PCR product is then purified and mixed with the linearized adenoviral vector of choice in the In-Fusion reaction. Following the reaction, a portion of the mixture is transformed into E. coli (Stellar Competent Cells) and screened. Once a PCR-positive clone is identified, the recombinant pAdenoX vector is amplified, purified, and subsequently linearized with the restriction enzyme PacI, then transfected into Adeno-X 293 cells for viral rescue and amplification. Adeno-X GoStix can be used to determine the status of adenovirus rescue.

Required Products

Cat. # Product Size Price License Quantity Details
631106 Tet System Approved FBS 500 mL $865.00

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

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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.

631101 Tet System Approved FBS, US-Sourced 500 mL $875.00

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.

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.

HEK 293 cells optimized for adenovirus production Adeno-X 293 cells
Adenovirus titration kits Adenovirus titration
Selection guides Adenovirus selection guide

Overview

  • Very tight control of gene expression
  • Simple-to-use, all-in-one tetracycline-inducible system
  • The most advanced adenoviral gene delivery technology
  • Easiest adenoviral system to use; cloning is even simpler than standard plasmid cloning

More 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.


Tech notes describing the use of adenoviral vector systems from Takara Bio

Fastest & easiest adenoviral vector system ever

Until now, the main drawback of commercially supplied adenoviral vector systems has been the need to use complex cloning procedures to overcome the difficulties with cloning into large (~34 kb) plasmids. Procedures have included precloning into shuttle vectors and subcloning through multiple steps and multiple different strains of E. coli, all of which increase hands-on time and leave more room for error. At Takara Bio, our Adeno-X virologists thought "wouldn't it be great if you could clone directly into the adenoviral plasmid just like any plasmid?" They then harnessed the power of In-Fusion HD Cloning technology to make this happen.

How it works Tet-inducible version

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FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2018 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.

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.

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED).

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