BSPII CRISPR Activation Plasmid (h): sc-402368-ACT

IBSP encodes bone sialoprotein II (BSPII), a secreted, highly acidic extracellular matrix glycoprotein enriched in mineralized tissues. BSPII binds hydroxyapatite and interacts with integrins through an RGD motif to support cell adhesion, migration, and osteoblast/osteoclast crosstalk during bone matrix deposition and remodeling. IBSP expression is regulated by osteogenic transcriptional programs and contributes to biomineralization-associated processes that intersect with focal adhesion and extracellular matrix organization pathways. Dysregulated IBSP/BSPII expression has been associated with altered bone turnover and is frequently used as a marker in studies of skeletal development, calcification, and tumor–bone microenvironment interactions.

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

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

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