Cytokeratin 10 CRISPR Activation Plasmid (h): sc-400193-ACT

KRT10 encodes cytokeratin 10, a type I intermediate filament protein that pairs with keratin 1 to form the suprabasal keratin network in differentiating epidermal keratinocytes. This cytoskeletal scaffold supports mechanical resilience, links to desmosomal adhesion, and coordinates terminal differentiation programs, including stratification and cornified envelope formation. KRT10 expression is tightly coupled to keratinocyte maturation and tissue-specific transcriptional control, and perturbations in keratin filament homeostasis can alter epithelial integrity and stress signaling. Genetic and expression changes involving KRT10 are implicated in disorders of epidermal differentiation and hyperkeratosis, providing a useful axis for studying skin barrier biology and epithelial disease mechanisms.

Cytokeratin 10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT10 expression without altering the underlying DNA sequence.

Cytokeratin 10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT10 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 KRT10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT10 locus and enabling the study of Cytokeratin 10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 10 pathway restoration in tumor cells with silenced or reduced KRT10 expression.

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