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EF1-alpha promoter versions of the Tet-On 3G inducible expression system

EF1alpha Tet-On 3G Systems

EF1-alpha promoter versions of the Tet-On 3G inducible expression system provide for consistent long-term expression of the Tet-On 3G transactivator, even in cell types known for their tendency to silence a CMV promoter over time, such as hematopoietic cells and stem cells. This means that in these cell types, long-term tetracycline-inducible expression can be achieved. 

EF1-alpha promoter versions of the Tet-On 3G inducible expression system provide for consistent long-term expression of the Tet-On 3G transactivator, even in cell types known for their tendency to silence a CMV promoter over time, such as hematopoietic cells and stem cells. This means that in these cell types, long-term tetracycline-inducible expression can be achieved. The systems contain the same pTRE3G vectors as our standard Tet-On 3G systems that generate the lowest possible background expression; however, the Tet-On 3G transactivator is expressed from an EF1-alpha promoter instead of a CMV promoter. The EF-1 alpha promoter in these systems is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1.

We tested the EF1-alpha version in Jurkat cells, a cell line known to show reduced expression and clonal variation in expression from CMV-based vectors. When expressing the Tet-On 3G transactivator protein from the EF1-alpha promoter, 83% of the Jurkat Tet-On 3G clones showed strong inducible expression and 33% demonstrated very high inducibility (greater than 2,000-fold). Such levels of control are not possible when using previous versions of the Tet-On system for this cell line.

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Cat. # Product Size Price License Quantity Details
631167 Tet-On® 3G Inducible Expression System (EF1alpha Version) Each Inquire for Quotation *

The Tet-On 3G Inducible Expression System is a powerful, tightly regulated, tetracycline-inducible mammalian expression system that is controlled by the addition of doxycycline to the culture medium. In the EF1α Version, Tet-On 3G transactivator expression is driven by a human elongation factor 1 alpha (EF1α) promoter, which allows expression of the protein in a variety of mammalian cells (such as hematopoietic or stem cells) without the transgene silencing associated with CMV promoters. The system includes a vector set, our highly efficient transfection reagent, Xfect,and Tet System Approved FBS.

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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Required Products

Cat. # Product Size Price License Quantity Details
631101 Tet System Approved FBS, US-Sourced 500 mL USD $971.00

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.

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631106 Tet System Approved FBS 500 mL USD $933.00

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.

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631342 Tet-On® 3G Bidirectional Inducible Expression System (EF1alpha, ZsGreen1) Each Inquire for Quotation *

The Tet-On 3G Inducible Expression System (EF1alpha, ZsGreen1) is a powerful, tightly regulated, tetracycline-inducible mammalian expression system that is controlled by the addition of doxycycline to the culture medium. This system allows the simultaneous expression of a gene of interest and a green fluorescent protein (ZsGreen1) marker from a bidirectional, Tet-responsive promoter that binds the Tet-On 3G transactivator protein in the presence of doxycycline. It includes the pTRE3G-BI-ZsGreen1 Vector Set, consisting of a bidirectional vector into which the gene of interest can be cloned and simultaneously expressed with the ZsGreen1 marker, a bidirectional control vector that contains the luciferase gene in one of the two possible cloning sites, and two linear selection markers for hygromycin and puromycin resistance—and the pEF1a-Tet3G Vector, which constitutively expresses the Tet-On 3G transactivator protein from a human elongation factor 1 alpha (EF1α) promoter, allowing expression of the protein in a variety of mammalian cells (such as hematopoietic or stem cells) without the transgene silencing associated with CMV promoters. This system also includes our highly efficient Xfect Transfection Reagent and Tet System Approved FBS.

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

Required Products

Cat. # Product Size Price License Quantity Details
631101 Tet System Approved FBS, US-Sourced 500 mL USD $971.00

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
631106 Tet System Approved FBS 500 mL USD $933.00

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
631347 Tet-On® 3G Inducible Expression System (EF1alpha, mCherry) Each Inquire for Quotation *

The Tet-On 3G Inducible Expression System (EF1alpha, mCherry) is a powerful, tightly regulated, tetracycline-inducible mammalian expression system that is controlled by the addition of doxycycline to the culture medium. This system allows the simultaneous expression of a gene of interest and a red fluorescent protein (mCherry) marker under the control of a Tet-responsive promoter that binds the Tet-On 3G transactivator protein in the presence of doxycycline. It includes the pTRE3G-mCherry Vector Set, consisting of an IRES-based vector into which the gene of interest can be cloned and simultaneously expressed with the mCherry marker, a control vector that inducibly expresses luciferase, and two linear selection markers for hygromycin and puromycin resistance—and the pEF1a-Tet3G Vector, which constitutively expresses the Tet-On 3G transactivator protein from a human elongation factor 1 alpha (EF1α) promoter, allowing expression of the protein in a variety of mammalian cells (such as hematopoietic or stem cells) without the transgene silencing associated with CMV promoters. This system also includes our highly efficient Xfect Transfection Reagent and Tet System Approved FBS.

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

Required Products

Cat. # Product Size Price License Quantity Details
631101 Tet System Approved FBS, US-Sourced 500 mL USD $971.00

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
631106 Tet System Approved FBS 500 mL USD $933.00

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
631363 Lenti-X™ Tet-On® 3G Inducible Expression System (EF1alpha Version) Each Inquire for Quotation *

The Lenti-X Tet-On 3G Inducible Expression System (EF1alpha Version) is a tetracycline-inducible lentiviral gene expression system that allows you to produce high titers of recombinant, VSV-G pseudotyped lentiviruses for the purpose of establishing a tightly inducible expression system in both dividing and non-dividing cell types. The pLVX-TRE3G Vector contains a TRE3G tetracycline-inducible promoter. The pLVX-EF1a-Tet3G Vector expresses the Tet-On 3G transactivator protein from the human EF1 alpha promoter, which allows for long-term expression of the transactivator in a broad range of cell types, including thosein which a CMV promoter can be silenced over time, such as hematopoietic stem cells. The kit also includes a complete fourth generation lentiviral packaging system,Lenti-X Packaging Single Shots (VSV-G).

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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Required Products

Cat. # Product Size Price License Quantity Details
631101 Tet System Approved FBS, US-Sourced 500 mL USD $971.00

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
631106 Tet System Approved FBS 500 mL USD $933.00

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

*You must be logged in to a Purchasing Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

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Overviews Tet-One overview

Overview

  • Tet-On 3G tetracyline transactivator is expressed from an EF1-alpha promoter
  • Very low background expression
  • High induced expression

More Information

Applications

  • Inducible expression in cell types known to silence CMV promoters, such as hematopoietic cells and stem cells

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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