ICAM-1/CD54 CRISPR Activation Plasmid (h): sc-400098-ACT

ICAM1 encodes intercellular adhesion molecule-1 (ICAM-1/CD54), an inducible immunoglobulin superfamily receptor expressed on endothelial and epithelial cells and many leukocytes. ICAM-1 binds β2 integrins such as LFA-1 (ITGAL/ITGB2) and Mac-1 (ITGAM/ITGB2) to mediate firm adhesion, transendothelial migration, and formation of immunological synapses. Its expression is strongly upregulated by inflammatory cues including TNF-α and IL-1 through NF-κB- and MAPK-dependent programs, linking ICAM1 to leukocyte trafficking and vascular inflammation. Dysregulated ICAM1 activity and expression are associated with chronic inflammatory states, autoimmune pathology, atherosclerotic processes, and tumor–immune interactions, making it a useful node for studying cell adhesion and cytokine-driven signaling.

ICAM-1/CD54 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ICAM1 expression without altering the underlying DNA sequence.

ICAM-1/CD54 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ICAM1 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 ICAM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ICAM-1/CD54 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ICAM1 locus and enabling the study of ICAM-1/CD54-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ICAM-1/CD54 pathway restoration in tumor cells with silenced or reduced ICAM1 expression.

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