Integrin β4/ITGB4/CD104 CRISPR Activation Plasmid (m): sc-431434-ACT

Itgb4 encodes integrin β4 (ITGB4/CD104), a transmembrane adhesion receptor that pairs primarily with integrin α6 to form α6β4, a key laminin receptor in epithelial basement membrane attachment. Through hemidesmosome assembly and crosstalk with focal adhesion and growth factor signaling, ITGB4 helps coordinate cell polarity, migration, and survival pathways including PI3K–AKT and MAPK signaling. Altered ITGB4 expression or signaling is associated with changes in epithelial integrity, invasive behavior, and extracellular matrix remodeling, making it relevant to studies of carcinoma progression, wound repair, and inflammatory tissue remodeling. In mouse systems, Itgb4 is frequently used to dissect epithelial–stromal interactions and adhesion-dependent signaling networks in organotypic and in vivo models.

Integrin β4/ITGB4/CD104 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Itgb4 expression without altering the underlying DNA sequence.

Integrin β4/ITGB4/CD104 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Itgb4 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 Itgb4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin β4/ITGB4/CD104 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Itgb4 locus and enabling the study of Integrin β4/ITGB4/CD104-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin β4/ITGB4/CD104 pathway restoration in tumor cells with silenced or reduced Itgb4 expression.

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