CMV-hspCas9-EF1-GFP SmartNuclease Pre-Packaged Lentiviral Particles [>10^6 IFUs]
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CMV-hspCas9-EF1-GFP SmartNuclease Pre-Packaged Lentiviral Particles [>10^6 IFUs]. Cat# CASLV105VA. Supplier: SBI System Biosciences
With SBI’s pCDH-CMV-Piranha™-EF1-GFP-T2A-Puro Pre-pacakged Lentivirus, you can build your own cell line constitutively expressing the Piranha Targeted Protein Degradation System. In this ready-to-transduce lentivirus, the Piranha protein is expressed from the strong CMV promoter and the construct is marked with GFP and puromycin. To initiate targeted protein degradation in your Piranha-expressing cell line, simply electroporate in antibody specific for your protein-of-interest—you’ll start seeing degradation of the protein-of-interest in as little as 1 hour*.
While studying protein function in vivo often involves knocking down expression using siRNA or knocking out the gene via CRISPR/Cas9, these approaches are indirect and often imperfect. siRNA can reduce or eliminate protein expression, but if the protein turn-over rate is slow then it can take days to see a reduction in function. Simply knocking out the gene coding for the protein-of-interest is not always possible, limits the type of studies you can do, and is incompatible with essential genes. To give researchers another faster option, SBI developed the Piranha Targeted Protein Degradation System.
- Specific—avoid off-target effects associated with genomic-based approaches (CRISPR/Cas9) or mRNA targeting (RNAi) by directly targeting the protein-of-interest
- Fast—observe protein degradation in as little as 1 hour* for speedier, more efficient studies
- Simple—relies on only two reagents, the Piranha System and your validated antibody
- Powerful—antibody-based targeting means the Piranha System can selectively degrade the phosphorylated or other post-translationally modified form of a protein while leaving the unmodified form intact
- Scalable for screening—use either the HEK293T Piranha Cell Line or your own cell line stably expressing Piranha protein to evaluate multiple protein targets in parallel (96- or 384-well plate assays)
- Unique—the only commercially available system for direct targeted degradation of protein(s)-of-interest for phenotypic studies
- Flexible—available as electroporation-ready mRNA, constitutively expressed in a HEK293T cell line, or as pre-packaged lentivirus for stable Piranha expression in the cell line of your choice
*Based on published data by Clift et al. A Method for the Acute and Rapid Degradation of Endogenous Proteins. Cell. 2017 Dec 14;171(7):1692-1706.e18. doi: 10.1016/j.cell.2017.10.033. Degradation rate of target protein and resulting phenotypic effects will depend on the specific protein being targeted and influenced by protein degradation pathway and any compensatory mechanisms. An initial time course experiment to find optimal conditions to assay for protein knockdown and phenotypes is highly recommended.Available Piranha Targeted Protein Degradation System Products Table 1. Available Piranha Targeted Protein Degradation System Products
How It Works
Targeted Protein Degradation with Piranha
SBI’s Piranha Targeted Protein Degradation System is based on the TRIM21 pathway found in most mammalian cells1 (Figure 1). Like TRIM21, the Piranha protein binds to conserved regions of an antibody, targeting the antibody—and the bound antigen—for degradation by the proteasome.
To use your newly-created Piranha-expressing cell line for targeted protein degradation, simply electroporate a validated antibody specific for the protein-of-interest into target cells, wait an appropriate amount of time, and then observe the phenotypic outcome. Because the rate of protein degradation is based upon many factors, we recommend performing initial experiments to empirically determine the degradation rate of your target protein and the optimal time for phenotypic observation.
1. Clift D, et al. A Method for the Acute and Rapid Degradation of Endogenous Proteins. Cell. 2017 Dec 14;171(7):1692-1706.e18. doi: 10.1016/j.cell.2017.10.033.
See the Piranha Targeted Protein Degradation System in action