Integrin α9/ITGA9 CRISPR Activation Plasmid (h): sc-402221-ACT

ITGA9 encodes integrin α9, an α subunit that heterodimerizes with integrin β1 to form a cell-surface receptor mediating adhesion to extracellular matrix ligands such as tenascin-C, osteopontin, and VCAM1. Integrin α9β1 signaling coordinates cytoskeletal remodeling, focal adhesion dynamics, and cell migration through pathways involving FAK/SRC, PI3K–AKT, and Rho family GTPases, thereby influencing tissue remodeling and inflammatory cell trafficking. ITGA9 activity has been linked to processes including angiogenesis, wound repair, and lymphatic valve development, and altered expression is studied in the context of fibrosis, chronic inflammation, and tumor cell invasiveness. As a mechanotransduction-associated receptor, integrin α9 provides a useful handle for interrogating how ECM composition and stiffness shape cellular behavior.

Integrin α9/ITGA9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGA9 expression without altering the underlying DNA sequence.

Integrin α9/ITGA9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGA9 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 ITGA9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin α9/ITGA9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGA9 locus and enabling the study of Integrin α9/ITGA9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin α9/ITGA9 pathway restoration in tumor cells with silenced or reduced ITGA9 expression.

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