Integrin β8/ITGB8 CRISPR Activation Plasmid (m): sc-435838-ACT

Itgb8 encodes integrin β8, a β subunit that pairs with αv to form the αvβ8 receptor, a cell-surface adhesion molecule that links extracellular matrix ligands to cytoskeletal and signaling networks. In mouse tissues, ITGB8 contributes to cell migration, tissue remodeling, and epithelial–mesenchymal communication, and it is a key regulator of latent TGF-β activation through interactions with ECM-associated complexes. Through crosstalk with focal adhesion signaling and TGF-β/SMAD pathways, ITGB8 influences differentiation programs, barrier properties, and immune cell behavior. Dysregulated ITGB8 activity has been associated with fibrotic remodeling, inflammatory microenvironments, and tumor biology in preclinical and translational studies, supporting its utility as a mechanistic node for pathway interrogation.

Integrin β8/ITGB8 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Itgb8 expression without altering the underlying DNA sequence.

Integrin β8/ITGB8 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Itgb8 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 Itgb8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin β8/ITGB8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Itgb8 locus and enabling the study of Integrin β8/ITGB8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin β8/ITGB8 pathway restoration in tumor cells with silenced or reduced Itgb8 expression.

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