periplakin CRISPR Activation Plasmid (h): sc-402350-ACT

Human PPL encodes periplakin, a plakin family cytolinker that connects intermediate filaments to desmosomes and the cornified envelope, supporting epithelial and endothelial mechanical stability. Periplakin participates in desmosome assembly, keratin network organization, and cytoskeletal remodeling during differentiation and wound responses, and it can influence signaling outputs linked to cell adhesion and stress pathways. Altered PPL expression or localization has been reported in multiple epithelial pathologies, including inflammatory skin disorders and cancers, where changes in cell–cell junction integrity and migratory behavior are relevant. As a scaffold at the cell periphery, periplakin is frequently studied in barrier formation, epithelial polarity, and junction-dependent regulation of proliferation and invasion.

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

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

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