Cytokeratin 12 CRISPR Activation Plasmid (h): sc-400955-ACT

KRT12 encodes cytokeratin 12, a type I intermediate filament protein that forms obligate heteropolymers with type II keratins to build the keratin cytoskeleton in differentiated corneal epithelium. This filament network supports epithelial mechanical integrity, shapes cell architecture, and coordinates junctional stability and cytoskeletal remodeling during stratification and wound repair–associated differentiation programs. KRT12 expression is tightly linked to corneal epithelial lineage identity and keratinization state, and its dysregulation is associated with corneal epithelial fragility and hereditary corneal dystrophy phenotypes. As a lineage-restricted structural marker, cytokeratin 12 is widely used to study epithelial differentiation, stress responses, and cytoskeleton-dependent signaling that integrates with cell adhesion and tissue homeostasis.

Cytokeratin 12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT12 expression without altering the underlying DNA sequence.

Cytokeratin 12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT12 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 KRT12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT12 locus and enabling the study of Cytokeratin 12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 12 pathway restoration in tumor cells with silenced or reduced KRT12 expression.

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