ESAM CRISPR Activation Plasmid (h): sc-413872-ACT

Endothelial cell-selective adhesion molecule (ESAM) is an immunoglobulin superfamily membrane protein enriched at endothelial tight junctions, where it supports cell–cell adhesion, vascular barrier integrity, and leukocyte transendothelial migration. ESAM participates in junctional organization and cytoskeletal regulation alongside other endothelial adhesion complexes, influencing angiogenic remodeling and inflammatory trafficking. Altered ESAM expression has been linked to vascular dysfunction, tumor-associated angiogenesis, and inflammatory disease contexts where endothelial permeability and immune cell infiltration are dysregulated. As a junctional marker and functional regulator, ESAM is commonly studied in endothelial differentiation, vascular permeability assays, and microenvironment-driven inflammation models.

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

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

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