osteopontin/OPN/SPP1 CRISPR Activation Plasmid (m): sc-423134-ACT

Mouse Spp1 encodes osteopontin (OPN/SPP1), a secreted phosphoglycoprotein that also associates with the extracellular matrix to regulate cell adhesion, migration, and survival. OPN signals through integrins and CD44 to modulate focal adhesion dynamics, cytoskeletal remodeling, and downstream pathways such as PI3K–AKT, MAPK, and NF-κB, shaping inflammatory and tissue remodeling responses. It is produced by osteoblast lineage cells, macrophages, and other stromal/immune compartments, contributing to bone mineralization, immune cell recruitment, and fibrosis-associated matrix remodeling. Dysregulated Spp1 expression is frequently linked to inflammatory disorders, cancer invasion/metastasis biology, and skeletal and renal pathophysiology in mouse disease models.

osteopontin/OPN/SPP1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Spp1 expression without altering the underlying DNA sequence.

osteopontin/OPN/SPP1 CRISPR Activation Plasmid (m) 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.