plakophilin 2 CRISPR Activation Plasmid (h): sc-402990-ACT

PKP2 encodes plakophilin 2, an armadillo repeat protein that localizes to desmosomes and intercalated discs where it stabilizes cell–cell adhesion by linking desmosomal cadherins to cytoskeletal networks. Through interactions with desmoplakin, plakoglobin, and intermediate filaments, plakophilin 2 contributes to junctional integrity, mechanotransduction, and tissue architecture, particularly in mechanically stressed epithelia and cardiac myocytes. PKP2 function also interfaces with signaling programs that respond to cell adhesion status and cytoskeletal tension. Genetic and functional alterations in PKP2 have been associated with arrhythmogenic cardiomyopathy phenotypes and related defects in myocardial conduction and structural remodeling, supporting its utility as a target in cardiac junction biology studies.

plakophilin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PKP2 expression without altering the underlying DNA sequence.

plakophilin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PKP2 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 PKP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous plakophilin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PKP2 locus and enabling the study of plakophilin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of plakophilin 2 pathway restoration in tumor cells with silenced or reduced PKP2 expression.

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