epiplakin 1 CRISPR Activation Plasmid (h): sc-404271-ACT

EPPK1 encodes epiplakin 1, a large plakin family cytolinker that associates with keratin intermediate filaments and contributes to cytoskeletal organization and epithelial mechanical stability. By scaffolding filament networks, epiplakin influences processes such as cell adhesion, migration, and stress responses that are integral to epithelial homeostasis and barrier integrity. Altered cytoskeletal coupling and epithelial architecture are frequently implicated in tissue injury and remodeling, making EPPK1 a relevant marker for studying epithelial differentiation states and stress-induced reorganization. Its expression and filament-associated functions are therefore of interest in research areas spanning epithelial biology, inflammation-associated tissue damage, and tumor cell plasticity.

epiplakin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPPK1 expression without altering the underlying DNA sequence.

epiplakin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPPK1 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 EPPK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous epiplakin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPPK1 locus and enabling the study of epiplakin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of epiplakin 1 pathway restoration in tumor cells with silenced or reduced EPPK1 expression.

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