Cytokeratin 7 CRISPR Activation Plasmid (h): sc-400628-ACT

KRT7 encodes cytokeratin 7, a type II intermediate filament protein that assembles with type I keratins to form the keratin cytoskeleton in simple epithelia. Cytokeratin 7 contributes to epithelial cell structural integrity, polarity, and mechanical resilience, and its organization is remodeled during differentiation, wound response, and stress signaling. Altered KRT7 expression patterns are widely used to study epithelial lineage identity and cytoskeletal reprogramming in contexts such as carcinoma subtype characterization and epithelial–mesenchymal transition. Because keratin networks interface with junctional complexes and intracellular trafficking, KRT7 is also relevant for investigating epithelial barrier function and cytoskeleton-linked signaling dynamics.

Cytokeratin 7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KRT7 expression without altering the underlying DNA sequence.

Cytokeratin 7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KRT7 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 KRT7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cytokeratin 7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KRT7 locus and enabling the study of Cytokeratin 7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cytokeratin 7 pathway restoration in tumor cells with silenced or reduced KRT7 expression.

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