Cytokeratin 1 CRISPR Activation Plasmid (h): sc-400893-ACT

KRT1 encodes cytokeratin 1, a type II intermediate filament protein that heterodimerizes with keratin 10 to form the suprabasal keratin network in stratified epithelia. This filament system provides mechanical resilience, supports desmosome-associated cell–cell adhesion, and participates in keratinocyte differentiation and barrier formation programs. Cytokeratin 1 dynamics interface with cytoskeletal remodeling and stress-response signaling that influence epithelial integrity during wound repair and inflammation. Dysregulated KRT1 expression or keratin filament organization is linked to epidermal fragility phenotypes and is frequently examined in studies of hyperkeratosis, keratinization defects, and epithelial stress adaptation.

Cytokeratin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT1 expression without altering the underlying DNA sequence.

Cytokeratin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT1 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 KRT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT1 locus and enabling the study of Cytokeratin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 1 pathway restoration in tumor cells with silenced or reduced KRT1 expression.

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