Cytokeratin 6B CRISPR Activation Plasmid (h): sc-401651-ACT

KRT6B encodes cytokeratin 6B, a type II intermediate filament protein that heterodimerizes with type I keratins to form the keratin cytoskeleton in stratified epithelia. Cytokeratin 6B supports mechanical resilience, cytoskeletal organization, and epithelial remodeling processes linked to wound repair and stress responses, integrating with junctional complexes and signaling programs that coordinate differentiation and proliferation. Altered KRT6B expression is observed in hyperproliferative keratinization states and epithelial pathology, and it is frequently used as a marker of activated keratinocytes in skin and mucosal tissues. These features make KRT6B relevant for studying epithelial homeostasis, barrier function, and cytoskeleton-associated changes in disease-relevant models.

Cytokeratin 6B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT6B expression without altering the underlying DNA sequence.

Cytokeratin 6B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT6B 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 KRT6B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 6B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT6B locus and enabling the study of Cytokeratin 6B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 6B pathway restoration in tumor cells with silenced or reduced KRT6B expression.

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