plakophilin 3 CRISPR Activation Plasmid (h): sc-403614-ACT

PKP3 encodes plakophilin 3, an armadillo-repeat protein that localizes to desmosomes and supports epithelial cell–cell adhesion by stabilizing desmosomal cadherin–plakoglobin complexes and linking junctional assemblies to intermediate filaments. Through regulation of desmosome assembly and junctional remodeling, plakophilin 3 contributes to tissue integrity, polarity, and mechanotransduction pathways that coordinate cytoskeletal organization. Altered PKP3 expression or desmosomal dysfunction has been associated with epithelial barrier defects and cancer-related phenotypes, including changes in migration and invasion programs. These properties make PKP3 a useful node for studying junction biology, cytoskeletal signaling, and context-dependent transcriptional responses to adhesion cues.

plakophilin 3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PKP3 expression without altering the underlying DNA sequence.

plakophilin 3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PKP3 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 PKP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous plakophilin 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PKP3 locus and enabling the study of plakophilin 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of plakophilin 3 pathway restoration in tumor cells with silenced or reduced PKP3 expression.

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