DSPP CRISPR Activation Plasmid (h): sc-401195-ACT

Dentin sialophosphoprotein (DSPP) is a secreted extracellular matrix precursor that is proteolytically processed into dentin sialoprotein and dentin phosphoprotein, key regulators of dentin mineralization and hydroxyapatite nucleation. In odontoblasts and related craniofacial mesenchymal lineages, DSPP contributes to matrix organization, phosphate handling, and mineral deposition processes that coordinate tooth development and structural integrity. Altered DSPP expression or function is linked to heritable dentin defects, enamel–dentin interface abnormalities, and broader craniofacial mineralization phenotypes. Because DSPP integrates extracellular matrix biology with biomineralization pathways, it serves as a useful molecular handle for studying odontogenic differentiation, matrix maturation, and mineral deposition dynamics in human cell models.

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

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

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