PKC nu CRISPR Activation Plasmid (h): sc-403822-ACT

PRKD3 encodes protein kinase D3 (PKC nu), a serine/threonine kinase that functions downstream of diacylglycerol-dependent PKC signaling to regulate phosphorylation networks controlling cell proliferation, survival, migration, and vesicular trafficking. PKC nu participates in signal integration from GPCR and receptor tyrosine kinase inputs and contributes to MAPK/ERK and NF-κB–linked transcriptional responses, with additional roles in cytoskeletal remodeling and Golgi-to-plasma membrane transport. Altered PRKD3 activity has been associated with dysregulated growth factor signaling and invasive phenotypes in multiple cancer contexts, making it a useful node for interrogating oncogenic pathway rewiring. In human cell models, PRKD3 perturbation is commonly used to study kinase-driven control of stress responses, motility programs, and transcriptional regulation.

PKC nu CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKD3 expression without altering the underlying DNA sequence.

PKC nu CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKD3 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 PRKD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKC nu expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKD3 locus and enabling the study of PKC nu-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKC nu pathway restoration in tumor cells with silenced or reduced PRKD3 expression.

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