osteopontin/OPN/SPP1 CRISPR Activation Plasmid (h): sc-400219-ACT

SPP1 encodes osteopontin (OPN), a secreted phosphoglycoprotein that functions as an extracellular matrix-associated cytokine influencing cell adhesion, migration, and survival through integrin and CD44 receptor engagement. OPN regulates inflammatory signaling and tissue remodeling by coordinating leukocyte chemotaxis, macrophage polarization, and osteoclast activity, and it integrates cues from focal adhesion and cytoskeletal dynamics to shape cell-matrix interactions. SPP1 expression is frequently altered in contexts of chronic inflammation, fibrosis, cardiovascular remodeling, and tumor–immune microenvironment crosstalk, where it can modulate invasion, angiogenesis, and immune cell recruitment. These properties make SPP1 a useful node for studying extracellular signaling, immune regulation, and matrix-driven phenotypes in human cell models.

osteopontin/OPN/SPP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPP1 expression without altering the underlying DNA sequence.

osteopontin/OPN/SPP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPP1 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 SPP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous osteopontin/OPN/SPP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPP1 locus and enabling the study of osteopontin/OPN/SPP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of osteopontin/OPN/SPP1 pathway restoration in tumor cells with silenced or reduced SPP1 expression.

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