Dermatopontin CRISPR Activation Plasmid (h): sc-402924-ACT

Human DPT encodes dermatopontin, an extracellular matrix (ECM) protein that binds collagens and decorates fibrillar networks to influence matrix organization, cell–matrix adhesion, and biomechanical properties of connective tissues. Dermatopontin contributes to fibroblast behavior and ECM remodeling by modulating interactions among collagen fibrils, proteoglycans, and cell-surface receptors, supporting processes such as wound repair and stromal architecture. Altered DPT expression and ECM composition are frequently examined in contexts of fibrosis, tumor-associated stroma, and vascular or dermal remodeling where changes in collagen organization and cellular adhesion affect tissue function. As a matrix-associated regulator, DPT is also relevant to studies of TGF-β–driven remodeling and broader mechanotransduction-linked transcriptional programs.

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

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

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