BR-cadherin CRISPR Activation Plasmid (h): sc-401987-ACT

CDH12 encodes BR-cadherin, a classical cadherin enriched in the nervous system that mediates Ca2+-dependent homophilic cell–cell adhesion and supports tissue architecture. Through catenin binding and linkage to the actin cytoskeleton, BR-cadherin participates in adherens junction organization, cell polarity, and coordinated migration programs that shape neural development. Altered cadherin expression can perturb epithelial/neuronal adhesion dynamics and influence signaling cross-talk with pathways such as Wnt/β-catenin and Rho GTPase-regulated cytoskeletal remodeling. CDH12 dysregulation has been reported in studies of neurodevelopmental phenotypes and tumor biology, motivating investigation of how adhesion-state changes impact invasion, connectivity, and differentiation.

BR-cadherin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDH12 expression without altering the underlying DNA sequence.

BR-cadherin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDH12 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 CDH12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BR-cadherin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDH12 locus and enabling the study of BR-cadherin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BR-cadherin pathway restoration in tumor cells with silenced or reduced CDH12 expression.

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