K-cadherin CRISPR Activation Plasmid (h): sc-401455-ACT

CDH6 encodes K-cadherin, a classical cadherin that mediates Ca²⁺-dependent cell–cell adhesion through homophilic binding and coupling of its cytoplasmic tail to catenins and the actin cytoskeleton. By organizing adherens junctions, K-cadherin influences epithelial architecture, contact inhibition, and coordinated cell migration during development and tissue maintenance. CDH6 expression is prominent in renal and neural lineages and is often studied in the context of epithelial–mesenchymal dynamics and lineage specification. Dysregulated cadherin programs, including altered CDH6 expression, are associated with changes in adhesion and invasiveness observed in multiple disease models, making CDH6 a useful node for investigating junctional signaling and cell-state transitions.

K-cadherin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDH6 expression without altering the underlying DNA sequence.

K-cadherin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDH6 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 CDH6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous K-cadherin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDH6 locus and enabling the study of K-cadherin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of K-cadherin pathway restoration in tumor cells with silenced or reduced CDH6 expression.

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