desmoplakin I/II CRISPR Activation Plasmid (h): sc-400818-ACT

DSP encodes desmoplakin I/II, a core cytolinker of desmosomes that anchors intermediate filaments to desmosomal plaques to maintain epithelial and cardiac tissue integrity under mechanical stress. Through interactions with desmogleins, desmocollins, plakoglobin, and plakophilins, desmoplakin supports cell–cell adhesion, cytoskeletal organization, and junctional remodeling during differentiation and wound-associated re-epithelialization. Perturbation of DSP expression or desmosome stability is associated with defective barrier function and altered mechanotransduction, with relevance to cardiomyopathic and skin fragility phenotypes. These features make DSP a useful target for investigating junctional signaling, intermediate filament dynamics, and stress-responsive pathways that couple adhesion to tissue homeostasis.

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

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

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