Cytokeratin 5 CRISPR Activation Plasmid (h): sc-400671-ACT

KRT5 encodes cytokeratin 5, a type II intermediate filament protein that pairs with keratin 14 to form the basal keratin network in stratified epithelia. Cytokeratin 5 contributes to mechanical stability, cell–cell adhesion architecture, and epithelial stress responses, and it is dynamically regulated during keratinocyte differentiation, wound repair, and basal cell fate maintenance. Altered KRT5 expression or keratin filament organization is associated with epithelial fragility phenotypes and is frequently evaluated in studies of basal-like epithelial programs and tumor lineage states. As a structural hub, cytokeratin 5 interfaces functionally with cytoskeletal remodeling and adhesion-linked signaling pathways that influence proliferation and barrier integrity.

Cytokeratin 5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT5 expression without altering the underlying DNA sequence.

Cytokeratin 5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT5 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 KRT5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT5 locus and enabling the study of Cytokeratin 5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 5 pathway restoration in tumor cells with silenced or reduced KRT5 expression.

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