VCAM-1 CRISPR Activation Plasmid (h): sc-400135-ACT

VCAM1 encodes vascular cell adhesion molecule 1 (VCAM-1), an inducible immunoglobulin superfamily adhesion receptor expressed on activated endothelium that mediates firm leukocyte adhesion and transendothelial migration through binding to integrins such as VLA-4 (α4β1). Its expression is strongly upregulated by inflammatory cytokines via NF-κB and MAPK signaling, linking endothelial activation to immune cell trafficking, vascular permeability changes, and tissue infiltration. VCAM-1 participates in vascular inflammation programs that intersect with oxidative stress responses and cytokine/chemokine networks. Dysregulated VCAM1 expression is associated with chronic inflammatory states, atherosclerotic lesion development, and tumor–stroma immune interactions, making it a useful molecular readout for endothelial activation and leukocyte recruitment biology.

VCAM-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VCAM1 expression without altering the underlying DNA sequence.

VCAM-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VCAM1 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 VCAM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VCAM-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VCAM1 locus and enabling the study of VCAM-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VCAM-1 pathway restoration in tumor cells with silenced or reduced VCAM1 expression.

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