Integrin β5/ITGB5 CRISPR Activation Plasmid (h): sc-401017-ACT

ITGB5 encodes integrin β5, a transmembrane adhesion receptor that heterodimerizes with integrin αV to form αVβ5, linking extracellular matrix ligands to the actin cytoskeleton and initiating outside-in signaling. Integrin β5 regulates focal adhesion dynamics, cell migration, and survival through pathways that include FAK/SRC, PI3K–AKT, and MAPK signaling, with additional roles in endocytosis and crosstalk with TGF-β–driven programs. By modulating cell–matrix interactions and mechanotransduction, ITGB5 influences epithelial and endothelial behavior, immune cell trafficking, and remodeling of the tumor microenvironment. Dysregulated ITGB5 expression or signaling has been associated with invasion, angiogenesis, fibrosis-related remodeling, and metastatic phenotypes across multiple disease contexts.

Integrin β5/ITGB5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGB5 expression without altering the underlying DNA sequence.

Integrin β5/ITGB5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGB5 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 ITGB5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin β5/ITGB5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGB5 locus and enabling the study of Integrin β5/ITGB5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin β5/ITGB5 pathway restoration in tumor cells with silenced or reduced ITGB5 expression.

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