Home › Products › Gene function › ProteoTuner protein control systems › Viral systems

Gene function

ProteoTuner expression systems in lentiviral and retroviral formats

ProteoTuner inducible expression systems in lentiviral and retroviral formats

Lentiviral and retroviral ProteoTuner systems make it possible to investigate the function of a specific protein of interest directly—by rapidly changing the abundance of the protein itself. These systems utilize a ligand-dependent destabilization domain (DD), and the ligand Shield1 to reversibly stabilize and destabilize a DD-tagged protein of interest in a predictable and dose-dependent manner. Your protein of interest is fused to a DD tag and rapidly stabilized by adding the Shield1 ligand to the culture medium. Plasmid formats are also available.

Cat. #

Product

Size

License

Quantity

Details

632175

Lenti-X™ ProteoTuner™ Shield System N (w/ ZsGreen1)

Each

USD $740.00

License Statement

ID Number
55	cPPT Element. This product and its use are the subject to one or more of the following U.S. Pat. No. 8,093,042. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA, Inc. is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).
57	This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.
K11	This product and its use are the subject to one or more of the following U.S. Pat. 8,093,042 and foreign equivalents. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).

The Lenti-X ProteoTuner Shield System N (w/ ZsGreen1) provides a unique method for quickly and directly regulating the amount of your protein of interest present in a cell, combined with versatile lentiviral delivery. The system utilizes a ligand-dependent destabilization domain (DD), derived from a 12 kDa mutant of the FKBP protein, and a membrane permeable stabilizing ligand, Shield1. Once cloned into the pLVX-PTuner Green Vector, the protein of interest is expressed with an N-terminal DD-tag and the fluorescent protein ZsGreen1. In the presence of Shield1, the DD-tagged protein is stabilized and will accumulate inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15–30 minutes after the addition of Shield1. In the absence of Shield1, the DD-tagged protein of interest is unstable. Consequently, removal of Shield1 (by splitting the cells into medium without Shield1) allows for destabilization and rapid degradation of the protein of interest. The extent of stabilization via Shield1 directly correlates with the amount of the ligand in the medium, so it is possible to fine-tune the amount of protein of interest present in the cell by controlling the amount of the Shield1 ligand.

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.

Components

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
632189	Shield1	500 uL	USD $360.00
The Shield1 compound stabilizes a protein of interest tagged with the destabilization domain (DD), which is based on a mutated version of the FKBP protein. It is used to protect the DD fusion protein of interest from proteasomal degradation. The amount of Shield1 can be varied to tune the amount of stabilized protein of interest within the cell. Documents Components Image Data Back DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance. While DD-N (solid blue line) is suitable for N-terminal fusions and even C-terminal fusions for many proteins, DD-C (dashed blue line) is optimal for C-terminal fusions and for those proteins that do not tolerate N-terminal fusions. HEK 293 cells expressing the DD fusions noted in the figure legend were treated with varying concentrations of Shield1. 12 hr later, the amount of AcGPF1 stabilized by different concentrations of Shield1 was detected by fluorescence intensity using flow cytometry. DD-C at the C-terminus of AcGFP showed a lower basal level of protein in the absence of Shield1 (purple line; see inset figure). MFI = mean fluorescence intensity. Back Structure of Shield1 stabilization ligand Structure of Shield1 stabilization ligand. Back Ligand-dependent, targeted and reversible protein stabilization Ligand-dependent, targeted and reversible protein stabilization. A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present. Back Protein stabilty controlled using the ProteoTuner shield system Protein stabilty controlled using the ProteoTuner shield system. DsRed-Express was cloned into pRetroX-PTuner IRES and used to infect HeLa cells. The amount of DD-tagged DsRed-Express stabilized by different concentrations of Shield1 was detected via Western blot using the Living Colors DsRed Polyclonal Antibody. Lane 1: molecular weight marker. Lane 2: 1X loading buffer. Lane 3: untreated HeLa cells (no virus, no Shield1). Lane 4: HeLa cells infected with the DD-DsRed Express construct; no Shield1. Lanes 5–8: HeLa cells infected with the DD-DsRed-Express construct and treated with 50, 250, 500, and 1,000 nM Shield1 respectively. Lane 9: 1X loading buffer. Lane 10: HEK 293 DsRed-Express stable cell line. Back 632189: Shield1 Back Control of actin stability with the ProteoTuner system Control of actin stability with the ProteoTuner system. In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).

632173

Lenti-X™ ProteoTuner™ Shield System N

Each

USD $740.00

License Statement

ID Number
57	This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.
55	cPPT Element. This product and its use are the subject to one or more of the following U.S. Pat. No. 8,093,042. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA, Inc. is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).
K11	This product and its use are the subject to one or more of the following U.S. Pat. 8,093,042 and foreign equivalents. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).

The Lenti-X ProteoTuner Shield System N provides a unique method for quickly and directly regulating the amount of your protein of interest present in a cell, combined with versatile lentiviral delivery. The system utilizes a ligand-dependent destabilization domain (DD), derived from a 12 kDa mutant of the FKBP protein, and a membrane permeable stabilizing ligand, Shield1. Once cloned into the pLVX-PTuner Vector, the protein of interest is expressed with an N-terminal DD-tag and the antibiotic selection marker puromycin. In the presence of Shield1, the DD-tagged protein is stabilized and will accumulate inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15–30 minutes after the addition of Shield1. In the absence of Shield1, the DD-tagged protein of interest is unstable. Consequently, removal of Shield1 (by splitting the cells into medium without Shield1) allows for destabilization and rapid degradation of the protein of interest. The extent of stabilization via Shield1 directly correlates with the amount of the ligand in the medium, so it is possible to fine-tune the amount of protein of interest present in the cell by controlling the amount of the Shield1 ligand.

Notice to purchaser

Components

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
632189	Shield1	500 uL	USD $360.00
The Shield1 compound stabilizes a protein of interest tagged with the destabilization domain (DD), which is based on a mutated version of the FKBP protein. It is used to protect the DD fusion protein of interest from proteasomal degradation. The amount of Shield1 can be varied to tune the amount of stabilized protein of interest within the cell. Documents Components Image Data Back DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance. While DD-N (solid blue line) is suitable for N-terminal fusions and even C-terminal fusions for many proteins, DD-C (dashed blue line) is optimal for C-terminal fusions and for those proteins that do not tolerate N-terminal fusions. HEK 293 cells expressing the DD fusions noted in the figure legend were treated with varying concentrations of Shield1. 12 hr later, the amount of AcGPF1 stabilized by different concentrations of Shield1 was detected by fluorescence intensity using flow cytometry. DD-C at the C-terminus of AcGFP showed a lower basal level of protein in the absence of Shield1 (purple line; see inset figure). MFI = mean fluorescence intensity. Back Structure of Shield1 stabilization ligand Structure of Shield1 stabilization ligand. Back Ligand-dependent, targeted and reversible protein stabilization Ligand-dependent, targeted and reversible protein stabilization. A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present. Back Protein stabilty controlled using the ProteoTuner shield system Protein stabilty controlled using the ProteoTuner shield system. DsRed-Express was cloned into pRetroX-PTuner IRES and used to infect HeLa cells. The amount of DD-tagged DsRed-Express stabilized by different concentrations of Shield1 was detected via Western blot using the Living Colors DsRed Polyclonal Antibody. Lane 1: molecular weight marker. Lane 2: 1X loading buffer. Lane 3: untreated HeLa cells (no virus, no Shield1). Lane 4: HeLa cells infected with the DD-DsRed Express construct; no Shield1. Lanes 5–8: HeLa cells infected with the DD-DsRed-Express construct and treated with 50, 250, 500, and 1,000 nM Shield1 respectively. Lane 9: 1X loading buffer. Lane 10: HEK 293 DsRed-Express stable cell line. Back 632189: Shield1 Back Control of actin stability with the ProteoTuner system Control of actin stability with the ProteoTuner system. In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).

632171

Retro-X™ ProteoTuner™ Shield System N

Each

USD $740.00

License Statement

ID Number
57	This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.

Direct manipulation of the presence and absence of a specific protein of interest is a very powerful tool for analyzing protein function. The Retro-X ProteoTuner Shield System N uses a unique method to regulate the amount of protein of interest present in a cell, quickly and directly. It utilizes a ligand-dependent destabilization domain (DD) and its membrane permeable stabilizing ligand, Shield1. The DD is based on a 12 kDa mutant of the FKBP protein, which is expressed as a tag onto the N-terminus of your protein of interest cloned into the pRetroX-PTuner Vector. In the presence of Shield1, the DD-tagged protein is stabilized and will accumulate inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15–30 minutes after the addition of Shield1. However, in the absence of the protective ligand Shield1, the DD-tagged protein of interest is rapidly degraded. Removing the Shield1 (by splitting the cells into media without Shield1) allows for protein destabilization, causing a fast degradation of the protein of interest. The extent of stabilization via Shield1 directly correlates with the amount of the ligand in the medium, so it is possible to tune the amount of protein of interest present in the cell, by controlling the amount of the Shield1 ligand.

Notice to purchaser

Components

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
632189	Shield1	500 uL	USD $360.00
The Shield1 compound stabilizes a protein of interest tagged with the destabilization domain (DD), which is based on a mutated version of the FKBP protein. It is used to protect the DD fusion protein of interest from proteasomal degradation. The amount of Shield1 can be varied to tune the amount of stabilized protein of interest within the cell. Documents Components Image Data Back DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance. While DD-N (solid blue line) is suitable for N-terminal fusions and even C-terminal fusions for many proteins, DD-C (dashed blue line) is optimal for C-terminal fusions and for those proteins that do not tolerate N-terminal fusions. HEK 293 cells expressing the DD fusions noted in the figure legend were treated with varying concentrations of Shield1. 12 hr later, the amount of AcGPF1 stabilized by different concentrations of Shield1 was detected by fluorescence intensity using flow cytometry. DD-C at the C-terminus of AcGFP showed a lower basal level of protein in the absence of Shield1 (purple line; see inset figure). MFI = mean fluorescence intensity. Back Structure of Shield1 stabilization ligand Structure of Shield1 stabilization ligand. Back Ligand-dependent, targeted and reversible protein stabilization Ligand-dependent, targeted and reversible protein stabilization. A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present. Back Protein stabilty controlled using the ProteoTuner shield system Protein stabilty controlled using the ProteoTuner shield system. DsRed-Express was cloned into pRetroX-PTuner IRES and used to infect HeLa cells. The amount of DD-tagged DsRed-Express stabilized by different concentrations of Shield1 was detected via Western blot using the Living Colors DsRed Polyclonal Antibody. Lane 1: molecular weight marker. Lane 2: 1X loading buffer. Lane 3: untreated HeLa cells (no virus, no Shield1). Lane 4: HeLa cells infected with the DD-DsRed Express construct; no Shield1. Lanes 5–8: HeLa cells infected with the DD-DsRed-Express construct and treated with 50, 250, 500, and 1,000 nM Shield1 respectively. Lane 9: 1X loading buffer. Lane 10: HEK 293 DsRed-Express stable cell line. Back 632189: Shield1 Back Control of actin stability with the ProteoTuner system Control of actin stability with the ProteoTuner system. In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).

632167

Retro-X™ ProteoTuner™ Shield System N (w/ ZsGreen1)

Each

USD $740.00

License Statement

ID Number
57	This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.

Direct manipulation of the presence and absence of a specific protein of interest is a very powerful tool for analyzing protein function. The Retro-X ProteoTuner Shield System N (w/ ZsGreen1) uses a unique method to regulate the amount of protein of interest present in a cell, quickly and directly. It utilizes a ligand-dependent destabilization domain (DD) and its membrane permeable stabilizing ligand, Shield1. The DD is based on a 12 kDa mutant of the FKBP protein, which is expressed as a tag on the N-terminus of your protein of interest. In the presence of Shield1, the DD-tagged protein is stabilized and will accumulate inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15–30 minutes after the addition of Shield1. However, in the absence of the protective ligand Shield1, the DD-tagged protein of interest is rapidly degraded. Removing the Shield1 (by splitting the cells into media without Shield1) allows for protein destabilization, causing a fast degradation of the protein of interest. The process of stabilization via Shield1 is reversible, so it is possible to tune the amount of protein of interest present in the cell, by controlling the amount of the Shield1 ligand.

Notice to purchaser

Components

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
632189	Shield1	500 uL	USD $360.00
The Shield1 compound stabilizes a protein of interest tagged with the destabilization domain (DD), which is based on a mutated version of the FKBP protein. It is used to protect the DD fusion protein of interest from proteasomal degradation. The amount of Shield1 can be varied to tune the amount of stabilized protein of interest within the cell. Documents Components Image Data Back DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance. While DD-N (solid blue line) is suitable for N-terminal fusions and even C-terminal fusions for many proteins, DD-C (dashed blue line) is optimal for C-terminal fusions and for those proteins that do not tolerate N-terminal fusions. HEK 293 cells expressing the DD fusions noted in the figure legend were treated with varying concentrations of Shield1. 12 hr later, the amount of AcGPF1 stabilized by different concentrations of Shield1 was detected by fluorescence intensity using flow cytometry. DD-C at the C-terminus of AcGFP showed a lower basal level of protein in the absence of Shield1 (purple line; see inset figure). MFI = mean fluorescence intensity. Back Structure of Shield1 stabilization ligand Structure of Shield1 stabilization ligand. Back Ligand-dependent, targeted and reversible protein stabilization Ligand-dependent, targeted and reversible protein stabilization. A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present. Back Protein stabilty controlled using the ProteoTuner shield system Protein stabilty controlled using the ProteoTuner shield system. DsRed-Express was cloned into pRetroX-PTuner IRES and used to infect HeLa cells. The amount of DD-tagged DsRed-Express stabilized by different concentrations of Shield1 was detected via Western blot using the Living Colors DsRed Polyclonal Antibody. Lane 1: molecular weight marker. Lane 2: 1X loading buffer. Lane 3: untreated HeLa cells (no virus, no Shield1). Lane 4: HeLa cells infected with the DD-DsRed Express construct; no Shield1. Lanes 5–8: HeLa cells infected with the DD-DsRed-Express construct and treated with 50, 250, 500, and 1,000 nM Shield1 respectively. Lane 9: 1X loading buffer. Lane 10: HEK 293 DsRed-Express stable cell line. Back 632189: Shield1 Back Control of actin stability with the ProteoTuner system Control of actin stability with the ProteoTuner system. In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).

631074

Lenti-X™ ProteoTuner™ Shield System C

Each

USD $740.00

License Statement

ID Number
55	cPPT Element. This product and its use are the subject to one or more of the following U.S. Pat. No. 8,093,042. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA, Inc. is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).
57	This product is covered by U.S. Patent Nos. 8,173,792 and 9,487,787.
K11	This product and its use are the subject to one or more of the following U.S. Pat. 8,093,042 and foreign equivalents. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot disclose information, sell or otherwise transfer this product, its components or materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. If the buyer is not willing to accept the limitations of this limited use statement, Takara Bio USA is willing to accept return of the product with a full refund. For information on purchasing a license to the DNA-Flap technology for purposes other than research, contact the Transfer of Technology Office, Institut Pasteur, 28 rue du Docteur Roux, 75 724 Paris Cedex 15 (www.pasteur.fr).

The Lenti-X ProteoTuner Shield System C provides a unique method for quickly and directly regulating the amount of your protein of interest present in a cell, and combines it with versatile lentiviral delivery. The system utilizes a ligand-dependent destabilization domain (DD-C) and a membrane-permeant stabilizing ligand, Shield1, to quickly and directly regulate the amount of your protein of interest present in a cell or organism. First, your gene of interest is cloned into the pLVX-PTunerC Vector, and expressed as a fusion with the destabilization domain (DD-C). Then the Shield1 ligand is either added or withheld, depending on your experiment.

In the presence of Shield1, the DD-C-tagged protein is stabilized and accumulates inside the cell. This ligand-dependent stabilization occurs very quickly, and has been observed as soon as 15-30 minutes after the addition of Shield1. In the absence of Shield1, the DD-C-tagged protein of interest is unstable. Consequently, removal of Shield1 allows for active and controllable degradation of the protein of interest. The extent of stabilization via Shield1 can be adjusted, and directly correlates with the amount of the ligand in the medium. Therefore, you can tune the level of your protein of interest present in the cell, by controlling the amount of the Shield1 ligand.

Notice to purchaser

Components

Required Products

Cat. #	Product	Size	Price	License	Quantity	Details
632189	Shield1	500 uL	USD $360.00
The Shield1 compound stabilizes a protein of interest tagged with the destabilization domain (DD), which is based on a mutated version of the FKBP protein. It is used to protect the DD fusion protein of interest from proteasomal degradation. The amount of Shield1 can be varied to tune the amount of stabilized protein of interest within the cell. Documents Components Image Data Back DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance. While DD-N (solid blue line) is suitable for N-terminal fusions and even C-terminal fusions for many proteins, DD-C (dashed blue line) is optimal for C-terminal fusions and for those proteins that do not tolerate N-terminal fusions. HEK 293 cells expressing the DD fusions noted in the figure legend were treated with varying concentrations of Shield1. 12 hr later, the amount of AcGPF1 stabilized by different concentrations of Shield1 was detected by fluorescence intensity using flow cytometry. DD-C at the C-terminus of AcGFP showed a lower basal level of protein in the absence of Shield1 (purple line; see inset figure). MFI = mean fluorescence intensity. Back Structure of Shield1 stabilization ligand Structure of Shield1 stabilization ligand. Back Ligand-dependent, targeted and reversible protein stabilization Ligand-dependent, targeted and reversible protein stabilization. A small destabilization domain (DD; blue) is fused to a target protein of interest. The small membrane-permeable ligand Shield1 (red) binds to the DD and protects it from proteasomal degradation. Removal of Shield1 however, causes rapid degradation of the entire fusion protein. The default pathway for the ProteoTuner systems is degradation of the fusion protein unless Shield1 is present. Back Protein stabilty controlled using the ProteoTuner shield system Protein stabilty controlled using the ProteoTuner shield system. DsRed-Express was cloned into pRetroX-PTuner IRES and used to infect HeLa cells. The amount of DD-tagged DsRed-Express stabilized by different concentrations of Shield1 was detected via Western blot using the Living Colors DsRed Polyclonal Antibody. Lane 1: molecular weight marker. Lane 2: 1X loading buffer. Lane 3: untreated HeLa cells (no virus, no Shield1). Lane 4: HeLa cells infected with the DD-DsRed Express construct; no Shield1. Lanes 5–8: HeLa cells infected with the DD-DsRed-Express construct and treated with 50, 250, 500, and 1,000 nM Shield1 respectively. Lane 9: 1X loading buffer. Lane 10: HEK 293 DsRed-Express stable cell line. Back 632189: Shield1 Back Control of actin stability with the ProteoTuner system Control of actin stability with the ProteoTuner system. In the absence of Shield1, no DD-AcGFP1 is observed (Panel B) despite the presence of a normal actin filament network (Panel A). In the presence of Shield1, DD-AcGFP1 is stable and present in the actin network (Panel D).

Overview

Rapid kinetics: protein level changes in minutes allows accurate functional analysis.
Precise tuning: precise control of protein level by controlling the dose of Shield1.
Reversible control: "protein on" to "protein off" for convincing gene-function studies.
What you get: each kit is supplied with a plasmid vector and an aliquot of Shield1.
NOTE: Most of the proteins that we tested show a better destabilization profile when the DD tag is fused to the N-terminus of the protein of interest (N Systems). Specific DD tag mutants for C-terminal tagging are available as well (C System); however, they have a slightly reduced destabilization activity in the absence of the Shield1 ligand.
For retrovirus production, use in combination with our Retro-X Universal Packaging System.
For lentivirus production, use in combination with our Lenti-X Packaging Single Shots.

More Information

Applications

Protein function in pathways
Functional analysis of subunits of a protein complex
Functional analysis of essential proteins

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.

Control protein stability using a small molecule ligand, Shield1

Control protein stability with Shield1

ProteoTuner systems allow quick, predictable regulation of protein presence or absence in mammalian cells by acting directly on the protein, using a small, cell-permeable synthetic compound. When Shield1 is added to the medium, any fusion protein containing a DD domain can accumulate to detectable levels within 15–20 minutes. Conversely, upon Shield1 removal, the half-time for the protein's degradation can be as short as 30 minutes. This post-translational regulation offers many benefits, from speed and convenience to precise tuning and reversible control.

Technology explained Product selection guide

Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2022 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.

ProteoTuner expression systems in lentiviral and retroviral formats

Notice to purchaser

Schematic of lentiviral ProteoTuner vectors

632175: Lenti-X ProteoTuner Shield System N (w/ ZsGreen1)

DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance

Structure of Shield1 stabilization ligand

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

632189: Shield1

Control of actin stability with the ProteoTuner system

Notice to purchaser

Dose-dependent stabilization of DD-tagged AcGFP1 fluorescent protein

Control of actin stability with the ProteoTuner system

Schematic of lentiviral ProteoTuner vectors

632173: Lenti-X ProteoTuner Shield System N

DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance

Structure of Shield1 stabilization ligand

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

632189: Shield1

Control of actin stability with the ProteoTuner system

Notice to purchaser

Dose-dependent stabilization of DD-tagged AcGFP1 fluorescent protein

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance

Structure of Shield1 stabilization ligand

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

632189: Shield1

Control of actin stability with the ProteoTuner system

Notice to purchaser

DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance

Structure of Shield1 stabilization ligand

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

632189: Shield1

Control of actin stability with the ProteoTuner system

Notice to purchaser

Schematic of lentiviral ProteoTuner vectors

DD fusions exhibit predictable, dose-dependent protein stabilization by Shield1; using the DD optimal for your protein is crucial in getting the best performance

Structure of Shield1 stabilization ligand

Ligand-dependent, targeted and reversible protein stabilization

Protein stabilty controlled using the ProteoTuner shield system

632189: Shield1

Control of actin stability with the ProteoTuner system

Overview

More Information

Additional product information