Integrin β3/ITGB3/CD61 CRISPR Activation Plasmid (m): sc-421175-ACT

Itgb3 encodes integrin β3 (ITGB3/CD61), a β subunit that pairs with αIIb or αV to form integrin receptors that mediate cell–cell and cell–matrix adhesion. Through ligand engagement and bidirectional signaling, integrin β3 regulates focal adhesion assembly, actin cytoskeletal remodeling, migration, and survival pathways including FAK/Src, PI3K–AKT, and MAPK signaling. In mouse systems, Itgb3 is widely studied in platelet and megakaryocyte biology, angiogenesis, osteoclast function, and inflammatory cell trafficking, linking its dysregulation to thrombosis-related phenotypes, vascular remodeling, and tumor–stroma interactions. These roles make ITGB3 a useful node for interrogating adhesion-dependent signaling and mechanotransduction in development and disease-relevant models.

Integrin β3/ITGB3/CD61 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Itgb3 expression without altering the underlying DNA sequence.

Integrin β3/ITGB3/CD61 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Itgb3 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 Itgb3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin β3/ITGB3/CD61 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Itgb3 locus and enabling the study of Integrin β3/ITGB3/CD61-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin β3/ITGB3/CD61 pathway restoration in tumor cells with silenced or reduced Itgb3 expression.

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