Piranha™ Electroporation-ready RFP-tagged mRNA (10µg)

Quickly and directly study the role a protein plays in vivo with SBI’s Piranha Targeted Protein Degradation System, Electroporation-ready mRNA. After electroporating both the Piranha mRNA and an antibody specific for your protein-of-interest into target cells, you can start seeing degradation of the protein-of-interest in as little as 1 hour*. Piranha Electroporation-ready mRNA is available in untagged, RFP-tagged, or GFP-tagged versions (also available as Pre-packaged Lentivirus or HEK293T Stable Cell Line).

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.

Figure 5. The speed of the Piranha System is also evident using a luciferase-based activity assay. MDA-MB-231 breast cancer cells were co-electroporated with Piranha mRNA and anti-IκBα antibody, and IκBα activity measured using an NFκB luciferase reporter assay—lower IκBα levels result in higher NFκB luciferase reporter activity. Compared to reduction of IκBα activity by siRNA (dark gray bars), which peaks on day 2, the Piranha System shows peak reduction of IκBα activity on day 1 (blue bars). The negative control is shown as light gray bars.

Figure 6. The Piranha System can reduce levels of phosphorylated protein. Piranha HEK293T Cells were electroporated with either buffer only (NC), anti-phospho-AKT(S473) antibody, or an IgG control, and total AKT levels assessed via Western blot.  Within 5-hrs post-electroporation, both phosphorylated and total AKT protein levels are reduced.