PKC eta CRISPR Activation Plasmid (h): sc-402297-ACT

PRKCH encodes protein kinase C eta (PKCη), a novel PKC family serine/threonine kinase activated downstream of diacylglycerol-dependent signaling and integrated with phospholipid-mediated pathways. PKCη contributes to regulation of keratinocyte differentiation, epithelial cell growth control, cytoskeletal remodeling, and stress-responsive signaling, with context-dependent roles in MAPK and NF-κB–linked transcriptional programs. In immune and barrier tissues, PRKCH activity influences cellular activation states and inflammatory signaling outputs, making it relevant to mechanistic studies of dysregulated proliferation, differentiation, and inflammation-associated phenotypes. Altered PRKCH expression or signaling has been investigated in disease biology where PKC isoform balance affects cellular behavior, supporting its use as a functional node in pathway interrogation.

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

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

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