PKC epsilon CRISPR Activation Plasmid (h): sc-400358-ACT

PRKCE encodes protein kinase C epsilon (PKCε), a diacylglycerol-responsive, Ca2+-independent serine/threonine kinase that couples receptor signaling to phosphorylation programs controlling proliferation, survival, migration, and cytoskeletal dynamics. PKCε integrates with GPCR and receptor tyrosine kinase inputs to modulate MAPK/ERK, PI3K–AKT, NF-κB, and cytoskeletal remodeling pathways, influencing vesicle trafficking and cell polarity. In human tissues, altered PRKCE expression or activity has been associated with dysregulated signal transduction in cancer biology, cardiovascular and ischemia–reperfusion responses, and neuroinflammatory processes. These features make PRKCE a relevant node for studying context-dependent kinase signaling, pathway cross-talk, and stress-adaptive phenotypes.

PKC epsilon CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKCE expression without altering the underlying DNA sequence.

PKC epsilon CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKCE locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the PRKCE transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKC epsilon expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKCE locus and enabling the study of PKC epsilon-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKC epsilon pathway restoration in tumor cells with silenced or reduced PRKCE expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.