PKC alpha CRISPR Activation Plasmid (h): sc-400128-ACT

PRKCA encodes protein kinase C alpha (PKC alpha), a classical, calcium- and diacylglycerol-responsive serine/threonine kinase that transduces signals downstream of phospholipase C to regulate phosphorylation-dependent control of cell growth, differentiation, cytoskeletal remodeling, and vesicular trafficking. PKC alpha integrates receptor-mediated cues to modulate MAPK/ERK signaling, NF-κB activity, and feedback within PI3K–AKT networks, shaping transcriptional programs and cellular stress responses. Altered PRKCA expression or PKC alpha signaling has been linked to dysregulated proliferation, invasion, and inflammatory signaling across multiple disease-associated contexts, making it a widely used node for mechanistic pathway interrogation. In human model systems, PRKCA perturbation supports studies of signal transduction dynamics, kinase-substrate relationships, and context-dependent pathway crosstalk.

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

PKC alpha CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKCA 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 PRKCA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKC alpha expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKCA locus and enabling the study of PKC alpha-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKC alpha pathway restoration in tumor cells with silenced or reduced PRKCA expression.

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