Lenti-X ProteoTuner Shield System N (w/ ZsGreen1)
Lenti-X ProteoTuner Shield System N (w/ ZsGreen1)
Brand: Takara Bio.
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Lenti-X ProteoTuner Shield System N (w/ ZsGreen1)
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.
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.
Applications
- Protein function in pathways
- Functional analysis of subunits of a protein complex
- Functional analysis of essential proteins
Schematic of lentiviral ProteoTuner vectors
Schematic of lentiviral ProteoTuner vectors.
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Schematic of lentiviral ProteoTuner vectors
Schematic of lentiviral ProteoTuner vectors.
