Integrin α4/ITGA4/CD49d CRISPR Activation Plasmid (h): sc-401336-ACT

ITGA4 encodes integrin α4 (CD49d), an α subunit that pairs with β1 or β7 to form VLA-4 (α4β1) or α4β7 integrins that mediate cell–cell and cell–matrix adhesion. Through binding to VCAM1, fibronectin CS-1, and MAdCAM1, integrin α4 regulates leukocyte trafficking, firm adhesion, and transendothelial migration, integrating outside-in signaling with cytoskeletal remodeling. ITGA4-linked signaling intersects with focal adhesion, MAPK, and PI3K pathways to influence survival, activation, and migratory behavior in immune and stromal compartments. Altered ITGA4 expression or function is commonly examined in studies of inflammatory cell recruitment, autoimmune-associated tissue infiltration, and tumor–microenvironment interactions involving metastatic dissemination and immune cell positioning.

Integrin α4/ITGA4/CD49d CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGA4 expression without altering the underlying DNA sequence.

Integrin α4/ITGA4/CD49d CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGA4 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 ITGA4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin α4/ITGA4/CD49d expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGA4 locus and enabling the study of Integrin α4/ITGA4/CD49d-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin α4/ITGA4/CD49d pathway restoration in tumor cells with silenced or reduced ITGA4 expression.

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