M-cadherin CRISPR Activation Plasmid (h): sc-401232-ACT

CDH15 encodes M-cadherin, a calcium-dependent cell–cell adhesion receptor enriched in skeletal muscle lineage cells and involved in myoblast recognition, fusion, and maintenance of tissue architecture. Through homophilic adhesion and coupling to catenins, M-cadherin supports adherens junction assembly, cytoskeletal organization, and contact-dependent signaling that influences differentiation and regenerative responses. CDH15 activity intersects with pathways governing cell migration and membrane dynamics during myogenesis, and altered expression has been reported in contexts of muscle degeneration/regeneration and tumor cell adhesion phenotypes. These features make CDH15 a useful target for studying adhesion-dependent regulation of lineage commitment and tissue remodeling in human cellular models.

M-cadherin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDH15 expression without altering the underlying DNA sequence.

M-cadherin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDH15 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 CDH15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous M-cadherin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDH15 locus and enabling the study of M-cadherin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of M-cadherin pathway restoration in tumor cells with silenced or reduced CDH15 expression.

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