plakophilin 1 CRISPR Activation Plasmid (h): sc-403577-ACT

PKP1 encodes plakophilin 1, an armadillo-repeat protein that localizes to desmosomes and supports calcium-dependent cell–cell adhesion by stabilizing desmosomal cadherin–plakoglobin complexes and linking junctional plaques to intermediate filaments. This scaffolding activity helps maintain epithelial integrity, barrier function, and mechanical resilience in stratified tissues, with additional roles reported in cytoskeletal organization and junctional signaling. Altered PKP1 expression or junctional mislocalization can disrupt desmosome assembly and epithelial differentiation, contributing to skin fragility phenotypes and aberrant tissue architecture. As a desmosome-associated factor, PKP1 is widely studied in pathways governing cell adhesion, tissue morphogenesis, and epithelial stress responses.

plakophilin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PKP1 expression without altering the underlying DNA sequence.

plakophilin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PKP1 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 PKP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous plakophilin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PKP1 locus and enabling the study of plakophilin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of plakophilin 1 pathway restoration in tumor cells with silenced or reduced PKP1 expression.

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