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  • Constitutive promoter
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Home › Products › Gene function › Viral transduction › Lentivirus › Vector systems › EF-1 alpha promoter

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Lentiviral vector systems with an EF-1 alpha promoter

Lentivirus with EF1-alpha promoters

Lenti-X lentiviral vector systems with EF-1 alpha promoters allow you to achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells. The EF-1 alpha promoter, which offers a broad host range, is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1 (Uetsuki et al. 1989; Wan Kim et al. 1990).

Lenti-X lentiviral vector systems with EF-1 alpha promoters allow you to achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells. The EF-1 alpha promoter, which offers a broad host range, is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1 (Uetsuki et al. 1989; Wan Kim et al. 1990).

Options for lentiviral vectors with an EF-1 alpha promoter

The Lenti-X Expression System (EF1α Version) is a complete system that includes the pLVX-EF1α-IRES-Puro Vector, and our fourth-generation lentiviral packaging system. Alternatively, you can express your protein fused to a red (mCherry or DsRed-Monomer) or green (AcGFP1) fluorescent protein, or express your protein and a fluorescent protein separately from an internal ribosome entry site (IRES).

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Cat. # Product Size Price License Quantity Details
631990 pLVX-EF1alpha-DsRed-Monomer-N1 Vector 10 ug Inquire for Quotation *

pLVX-EF1α-DsRed-Monomer-N1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the N-terminus of the fluorescent protein DsRed-Monomer. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells), without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing DsRed-Monomer fusions, without time-consuming drug and clonal selection.

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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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631989 pLVX-EF1alpha-DsRed-Monomer-C1 Vector 10 ug Inquire for Quotation *

pLVX-EF1α-DsRed-Monomer-C1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the C-terminus of the fluorescent protein DsRed-Monomer. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing DsRed-Monomer fusions, without time-consuming drug and clonal selection.

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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631989: pLVX-EF1alpha-DsRed-Monomer-C1 Vector

631989: pLVX-EF1alpha-DsRed-Monomer-C1 Vector

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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631987 pLVX-EF1alpha-IRES-mCherry Vector 10 ug Inquire for Quotation *

pLVX-EF1α-IRES-mCherry is a bicistronic lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector contains an internal ribosomal entry site (IRES) that allows a gene-of-interest and the red fluorescent protein mCherry to be simultaneously coexpressed from a single mRNA transcript. Expression of the transcript is driven by the constitutively active human elongation factor 1 alpha (EF1α) promoter.

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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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

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631987: pLVX-EF1alpha-IRES-mCherry Vector

631987: pLVX-EF1alpha-IRES-mCherry Vector

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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

Required Products

Cat. # Product Size Price License Quantity Details
631275 Lenti-X™ Packaging Single Shots (VSV-G) 16 Rxns USD $1071.00

License Statement

ID Number  
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.: 8,562,966, 8,557,231. 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 one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

Lenti-X Packaging Single Shots (VSV-G) provide an extremely simple and consistent one-step method for producing high-titer lentivirus. No additional transfection reagent is needed because Lenti-X Packaging Single Shots (VSV-G) consist of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral vector of choice in sterile water and adding it to 293T cells, e.g., Lenti-X 293T Cells (Cat. # 632180), in a 10 cm dish.

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The Lenti-X Packaging Single Shots (VSV-G) protocol

The Lenti-X Packaging Single Shots (VSV-G) protocol
The Lenti-X Packaging Single Shots (VSV-G) protocol.

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Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers. A lentiviral vector containing the ZsGreen1 gene was packaged according to the Lenti-X single shots protocol in four independent experiments. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots; the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. 48 hours after transfection, the cells were imaged by fluorescence microscopy (Panel A, top) and light microscopy (Panel A, bottom). After images were taken, the supernatant was harvested and used infect HT1080 cells for titer determination (Panel B, IFU/ml).

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High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots. A CMV ZsGreen1 expression cassette was cloned into several lentiviral vector backbones. These vectors were then packaged into lentivirus using the Lenti-X packaging single shots following the provided protocol. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots, and the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. After 48 hours, titer was determined using several methods. To determine infectivity, the supernatant was harvested and used to infect HT1080 cells (Flow Cytometry). Harvested viral supernatants were also analyzed by RT-PCR to quantify viral genome copies (qRT-PCR, Lenti-X qRT-PCR Titration Kit), ELISA to measure p24 (p24 ELISA, Lenti-X p24 Rapid Titer Kit), and by a rapid lentiviral detection method (Lenti-X GoStix).

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A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems . Our Lenti-X Packaging single shots utilize a packaging system that consists of five separate components (Panel A), mixed in proprietary proportions for optimized packaging activity. The separation of the gag, pol, and env genes effectively reduces the incidence of RCL (Wu et al., 2000). High levels of expression of essential viral components are driven by the Tet-Off and Tat transactivators, which induce a cascade of expression that results in high titers of lentivirus. The pol gene is fused to vpr to ensure transport of the reverse transcriptase/integrase protein into the recombinant lentiviral particle. Not all vector elements are shown. Other 3rd generation lentiviral packaging systems (Panel B) generate lower titers, do not contain separate gag and pol sequences, and do not use a transactivation cascade mechanism.

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631275: Lenti-X Packaging Single Shots (VSV-G)

631275: Lenti-X Packaging Single Shots (VSV-G)
632180 Lenti-X™ 293T Cell Line 1 mL USD $422.00

License Statement

ID Number  
406 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The Lenti-X 293T Cell Line is a subclone of the transformed human embryonic kidney cell line, HEK 293, which is highly transfectable and supports high levels of viral protein expression. When transfected with Lenti-X Packaging Single Shots and a lentiviral vector, these cells are capable of producing lentiviral titers as high as >108 ifu/ml, as determined by flow cytometry. The cell line also constitutively expresses the simian virus 40 (SV40) large T antigen.

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632180: Lenti-X 293T Cell Line

632180: Lenti-X 293T Cell Line

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Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein

Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein
Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein. As little as 10 µl of supernatant from Lenti-X transduced the majority of these HeLa cells, whereas 10 µl of supernatant from the other system transduced only a small percentage of the cells. Transduced cells were quantified by flow cytometry.

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Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus. Recombinant lentivirus for expressing ZsGreen1 was produced using Lenti-X virus and used to transduce normal human neural progenitor cells. A single transduced cell is shown under phase contrast microscopy (Panel A) and fluorescence microscopy (Panel B).

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High-titer lentivirus production

High-titer lentivirus production

High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.

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293T cell line for higher titers

293T cell line for higher titers

293T cell line for higher titers. We used our fourth-generation lentiviral packaging system and one of our pLVX-lentiviral vectors to compare the virus production of the Lenti-X 293T Cell Line to that of two other commonly used HEK 293-based cell lines. Lenti-X 293T cells clearly outperformed the other cell lines—producing over 6X more virus than 293FT cells and up to 30X more virus than the parental HEK 293 cell line.

631986 pLVX-EF1alpha-mCherry-N1 Vector 10 ug Inquire for Quotation *

pLVX-EF1α-mCherry-N1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the N-terminus of the red fluorescent protein mCherry. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing mCherry fusions, without time-consuming drug and clonal selection.

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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631986: pLVX-EF1alpha-mCherry-N1 Vector

631986: pLVX-EF1alpha-mCherry-N1 Vector

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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

Back

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631985 pLVX-EF1alpha-mCherry-C1 Vector 10 ug Inquire for Quotation *

pLVX-EF1α-mCherry-C1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the C-terminus of the red fluorescent protein mCherry. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing mCherry fusions, without time-consuming drug and clonal selection.

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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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

Back

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631984 pLVX-EF1alpha-AcGFP1-C1 Vector 10 ug Inquire for Quotation

License Statement

ID Number  
39 AcGFP is covered by U.S. Patent Numbers; 7,432,053, 7,667,016, 7,879,988 and 7,897,726.
72 Living Colors Fluorescent Protein Products: Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such license specifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc. Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com. For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by e-mail at licensing@takarabio.com. Not-For-Profit Non-Commercial Use License: A copy of the pLVX-EF1alpha-AcGFP1-C1 Vector product License Agreement can be found by clicking here.
*

pLVX-EF1α-AcGFP1-C1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the C-terminus of the green fluorescent protein AcGFP1. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing AcGFP1 fusions, without time-consuming drug and clonal selection.

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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631984: pLVX-EF1alpha-AcGFP1-C1 Vector

631984: pLVX-EF1alpha-AcGFP1-C1 Vector

Back

Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

Back

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631983 pLVX-EF1alpha-AcGFP1-N1 Vector 10 ug Inquire for Quotation

License Statement

ID Number  
39 AcGFP is covered by U.S. Patent Numbers; 7,432,053, 7,667,016, 7,879,988 and 7,897,726.
72 Living Colors Fluorescent Protein Products: Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such license specifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc. Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at licensing@takarabio.com. For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by e-mail at licensing@takarabio.com. Not-For-Profit Non-Commercial Use License: A copy of the pLVX-EF1alpha-AcGFP1-N1 Vector product License Agreement can be found by clicking here.
*

pLVX-EF1α-AcGFP1-N1 is a lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector allows a gene-of-interest to be fused to the N-terminus of the green fluorescent protein AcGFP1. Expression of the fusion is driven by the human elongation factor 1 alpha (EF1α) promoter, which continues to be constitutively active even after stable integration of the vector into the host cell genome. Stable expression of the fusion allows the monitoring of a variety of cellular processes (such as differentiation in primary or stem cells) without the transgene silencing associated with CMV promoters. In addition, the vector allows efficient flow cytometric detection of stably or transiently transfected mammalian cells expressing AcGFP1 fusions, without time-consuming drug and clonal selection.

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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631983: pLVX-EF1alpha-AcGFP1-N1 Vector

631983: pLVX-EF1alpha-AcGFP1-N1 Vector

Back

Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

Back

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

631982 pLVX-EF1alpha-IRES-ZsGreen1 Vector 10 ug USD $705.00

pLVX-EF1α-IRES-ZsGreen1 is a bicistronic lentiviral expression vector that can be used to generate high-titer lentivirus for transducing virtually any dividing or nondividing mammalian cell type, including primary and stem cells. The vector contains an internal ribosomal entry site (IRES) that allows a gene-of-interest and the ZsGreen1 fluorescent protein to be simultaneously coexpressed from a single mRNA transcript. Expression of the transcript is driven by the constitutively active human elongation factor 1 alpha (EF1α) promoter.

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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631982: pLVX-EF1alpha-IRES-ZsGreen1 Vector

631982: pLVX-EF1alpha-IRES-ZsGreen1 Vector

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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

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Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter

Expression of AcGFP1 driven by the EF-1 alpha promoter in stem cell lines is higher than expression driven by the CMV promoter. The mouse embryonic stem cell lines E14 (Panel A) and D3 (Panel B) were transduced by Lenti-X lentivirus, expressing AcGFP1 either under the control of the CMV promoter or the Elongation factor alpha (EF-1 alpha) promoter. The expression level of AcGFP1 in infected cells five days postinfection was monitored by FACS analysis using the FL1 channel. The expression of AcGFP1 driven by the EF-1 alpha promoter in both stem cell lines was considerably higher compared to the CMV promoter. This is mainly due to a considerably lower rate of silencing of the EF-1 alpha promoter in stem cells compared to the CMV promoter as published (Wang, et al. (2008) Stem Cells Dev 17:279–289).

Required Products

Cat. # Product Size Price License Quantity Details
631275 Lenti-X™ Packaging Single Shots (VSV-G) 16 Rxns USD $1071.00

License Statement

ID Number  
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.: 8,562,966, 8,557,231. 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 one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

Lenti-X Packaging Single Shots (VSV-G) provide an extremely simple and consistent one-step method for producing high-titer lentivirus. No additional transfection reagent is needed because Lenti-X Packaging Single Shots (VSV-G) consist of pre-aliquoted, lyophilized, single tubes of Xfect Transfection Reagent premixed with an optimized formulation of VSV-G pseudotyped Lenti-X lentiviral packaging plasmids. High-titer virus is produced by simply reconstituting this mixture with your lentiviral vector of choice in sterile water and adding it to 293T cells, e.g., Lenti-X 293T Cells (Cat. # 632180), in a 10 cm dish.

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The Lenti-X Packaging Single Shots (VSV-G) protocol

The Lenti-X Packaging Single Shots (VSV-G) protocol
The Lenti-X Packaging Single Shots (VSV-G) protocol.

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Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers

Consistent, high-efficiency transfections lead to high titers. A lentiviral vector containing the ZsGreen1 gene was packaged according to the Lenti-X single shots protocol in four independent experiments. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots; the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. 48 hours after transfection, the cells were imaged by fluorescence microscopy (Panel A, top) and light microscopy (Panel A, bottom). After images were taken, the supernatant was harvested and used infect HT1080 cells for titer determination (Panel B, IFU/ml).

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High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots

High-titer virus was produced regardless of the lentiviral vector backbone with Lenti-X packaging single shots. A CMV ZsGreen1 expression cassette was cloned into several lentiviral vector backbones. These vectors were then packaged into lentivirus using the Lenti-X packaging single shots following the provided protocol. Briefly, 7 µg of pLVX-ZsGreen1 plasmid was added to each of four Lenti-X single shots, and the tubes were vortexed for 20 sec and incubated at room temperature for 10 min. Then, the mixture was added to cultured Lenti-X 293T cells that were approximately 80% confluent. After 48 hours, titer was determined using several methods. To determine infectivity, the supernatant was harvested and used to infect HT1080 cells (Flow Cytometry). Harvested viral supernatants were also analyzed by RT-PCR to quantify viral genome copies (qRT-PCR, Lenti-X qRT-PCR Titration Kit), ELISA to measure p24 (p24 ELISA, Lenti-X p24 Rapid Titer Kit), and by a rapid lentiviral detection method (Lenti-X GoStix).

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A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems

A comparison of fourth- and third-generation lentiviral packaging systems . Our Lenti-X Packaging single shots utilize a packaging system that consists of five separate components (Panel A), mixed in proprietary proportions for optimized packaging activity. The separation of the gag, pol, and env genes effectively reduces the incidence of RCL (Wu et al., 2000). High levels of expression of essential viral components are driven by the Tet-Off and Tat transactivators, which induce a cascade of expression that results in high titers of lentivirus. The pol gene is fused to vpr to ensure transport of the reverse transcriptase/integrase protein into the recombinant lentiviral particle. Not all vector elements are shown. Other 3rd generation lentiviral packaging systems (Panel B) generate lower titers, do not contain separate gag and pol sequences, and do not use a transactivation cascade mechanism.

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631275: Lenti-X Packaging Single Shots (VSV-G)

631275: Lenti-X Packaging Single Shots (VSV-G)
632180 Lenti-X™ 293T Cell Line 1 mL USD $422.00

License Statement

ID Number  
406 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The Lenti-X 293T Cell Line is a subclone of the transformed human embryonic kidney cell line, HEK 293, which is highly transfectable and supports high levels of viral protein expression. When transfected with Lenti-X Packaging Single Shots and a lentiviral vector, these cells are capable of producing lentiviral titers as high as >108 ifu/ml, as determined by flow cytometry. The cell line also constitutively expresses the simian virus 40 (SV40) large T antigen.

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632180: Lenti-X 293T Cell Line

632180: Lenti-X 293T Cell Line

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Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein

Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein
Clontech's lentiviral packaging system (Panel A) and a lentiviral packaging system from a leading competitor (Panel B) were each used to generate viral supernatants from their respective lentiviral system vectors that were engineered to express the ZsGreen1 fluorescent protein. As little as 10 µl of supernatant from Lenti-X transduced the majority of these HeLa cells, whereas 10 µl of supernatant from the other system transduced only a small percentage of the cells. Transduced cells were quantified by flow cytometry.

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Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus

Transduction of neural progenitor cells by Lenti-X lentivirus. Recombinant lentivirus for expressing ZsGreen1 was produced using Lenti-X virus and used to transduce normal human neural progenitor cells. A single transduced cell is shown under phase contrast microscopy (Panel A) and fluorescence microscopy (Panel B).

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High-titer lentivirus production

High-titer lentivirus production

High-titer lentivirus production. Lenti-X 293T cells were transduced with the indicated volumes (µl) of lentiviral packaging supernatant generated with the Lenti-X Expression System and then selected with puromycin for 9 days to allow the formation of the resistant colonies, which were then stained with crystal violet.

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293T cell line for higher titers

293T cell line for higher titers

293T cell line for higher titers. We used our fourth-generation lentiviral packaging system and one of our pLVX-lentiviral vectors to compare the virus production of the Lenti-X 293T Cell Line to that of two other commonly used HEK 293-based cell lines. Lenti-X 293T cells clearly outperformed the other cell lines—producing over 6X more virus than 293FT cells and up to 30X more virus than the parental HEK 293 cell line.

631253 Lenti-X™ Expression System (EF1alpha Version) Each USD $1140.00

License Statement

ID Number  
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.: 8,562,966, 8,557,231. 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.
89
This Product is protected by one or more patents from the family consisting of: US9945850 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.

The Lenti-X Expression System (EF1α Version) enables you to produce exceptionally high titers of recombinant lentivirus for expression of any cDNA in any cell type susceptible to lentiviral transduction. The System contains an HIV-1-based, lentiviral expression vector, pLVX-EF1α-IRES-Puro, designed to constitutively express a protein of interest and puromycin resistance (Puror). Constitutive expression of these proteins is driven by the human elongation factor 1 alpha (EF1α) promoter, which allows protein expression in mammalian cells in which the CMV promoter is silenced. The System also includes Lenti-X Packaging Single Shots (VSV-G) and Lenti-X GoStix.

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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4th generation lentiviral packaging system

4th generation lentiviral packaging system

Fourth-generation lentiviral packaging system. A lentiviral vector and the Lenti-X HTX Packaging Mix are cotransfected into 293T cells. High titer lentiviral supernatants are ready for use 48 hr after transfection.

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Lentiviral vector systems with EF-1 alpha promoters

Lentiviral vector systems with EF-1 alpha promoters
Lentiviral vector systems with EF-1 alpha promoters. Achieve robust, constitutive, long-term expression of your gene of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells.

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631253: Lenti-X Expression System (EF1alpha Version)

631253: Lenti-X Expression System (EF1alpha Version)

*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.

Selection guides Lentiviral vector options
Videos About Lenti-X packaging
Overviews Premade lentiviral particles

Overview

  • High-titer lentiviral vector system
  • Constitutive expression from the human EF1 alpha promoter
  • Robust, long-term expression, in the broadest range of cell types
  • Bright fluorescent proteins to monitor expression or localization, learn more from our fluorescent protein quick guide.

More Information

Applications

  • If CMV promoters are silenced in your cell type, try EF-1 alpha
  • Lentiviral transduction of stem cells
  • Lentiviral transduction of hematopoietic cells
References

Uetsuki, T., Naito, A., Nagata, S. & Kaziro, Y. Isolation and characterization of the human chromosomal gene for polypeptide chain elongation factor-1. J. Biol. Chem. 264, 5791–5798 (1989).

Wan Kim, D., Uetsuki, T., Kaziro, Y., Yamaguchi, N. & Sugano, S. Use of the human alongation factor 1a promoter as a versatile and efficient expression system. Gene 91, 217–223 (1990).

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.


Lentiviral workflow

Lentiviral workflow

Find learning resources and technical information matched to each step of your viral transduction workflow.

Lentiviral workflow Transduction learning center

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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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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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