PCDH8 CRISPR Activation Plasmid (h): sc-405324-ACT

PCDH8 (protocadherin 8) is a calcium-dependent cell–cell adhesion molecule of the cadherin superfamily that contributes to tissue architecture and intercellular signaling in human cells. By modulating adhesion dynamics and contact-dependent communication, PCDH8 influences processes such as cellular polarity, migration, and differentiation, with downstream effects on cytoskeletal organization and junctional remodeling. Dysregulated protocadherin expression patterns have been associated with altered adhesion and invasive phenotypes in cancer biology, and PCDH8 is frequently investigated in the context of epigenetic regulation and transcriptional control. As part of broader cadherin/protocadherin networks, PCDH8 provides a tractable node for dissecting mechanisms that couple cell-surface adhesion to gene regulatory programs.

PCDH8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCDH8 expression without altering the underlying DNA sequence.

PCDH8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCDH8 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 PCDH8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PCDH8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCDH8 locus and enabling the study of PCDH8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PCDH8 pathway restoration in tumor cells with silenced or reduced PCDH8 expression.

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