Integrin β6/ITGB6 CRISPR Activation Plasmid (h): sc-400999-ACT

ITGB6 encodes the integrin β6 subunit, which pairs with integrin αV to form the αVβ6 heterodimer that mediates epithelial cell adhesion to extracellular matrix ligands and regulates migration, polarity, and tissue remodeling. αVβ6 is a context-dependent activator of latent TGF-β through interactions with RGD-containing proteins, linking ITGB6 to TGF-β/SMAD signaling, epithelial–mesenchymal transition, and wound repair programs. Aberrant ITGB6 expression and αVβ6 function have been associated with fibrotic remodeling and tumor-associated invasion and metastasis in multiple epithelial cancers, where it can modulate the local microenvironment and inflammatory signaling. As a surface receptor integrating ECM cues with intracellular signaling, ITGB6 is frequently used to study mechanotransduction, cell–matrix communication, and TGF-β–driven transcriptional responses.

Integrin β6/ITGB6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGB6 expression without altering the underlying DNA sequence.

Integrin β6/ITGB6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGB6 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 ITGB6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin β6/ITGB6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGB6 locus and enabling the study of Integrin β6/ITGB6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin β6/ITGB6 pathway restoration in tumor cells with silenced or reduced ITGB6 expression.

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