Integrin α8/ITGA8 CRISPR Activation Plasmid (h): sc-402020-ACT

ITGA8 encodes integrin α8, an α subunit that heterodimerizes with β1 to form a laminin- and extracellular matrix–binding receptor that regulates cell–matrix adhesion, cytoskeletal organization, and mechanotransduction. Integrin α8/ITGA8 contributes to focal adhesion signaling and downstream pathways involving FAK/SRC, Rho-family GTPases, and MAPK that coordinate cell migration, differentiation, and tissue remodeling. It is prominently studied in mesenchymal and stromal contexts, including kidney development and fibroblast biology, where altered integrin-mediated signaling can influence extracellular matrix deposition and organ morphogenesis. Dysregulated ITGA8 expression or integrin signaling has been associated with fibrotic phenotypes and tumor microenvironment remodeling, making it relevant for investigations of invasion, adhesion-dependent survival, and matrix-driven transcriptional programs.

Integrin α8/ITGA8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGA8 expression without altering the underlying DNA sequence.

Integrin α8/ITGA8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGA8 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 ITGA8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin α8/ITGA8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGA8 locus and enabling the study of Integrin α8/ITGA8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin α8/ITGA8 pathway restoration in tumor cells with silenced or reduced ITGA8 expression.

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