MAdCAM-1 CRISPR Activation Plasmid (h): sc-403616-ACT

MADCAM1 encodes mucosal addressin cell adhesion molecule 1 (MAdCAM-1), an endothelial glycoprotein that mediates selective recruitment and retention of circulating lymphocytes within mucosal tissues. By engaging the α4β7 integrin on lymphocytes, MAdCAM-1 supports adhesion and transmigration across specialized venules and contributes to the organization of gut-associated lymphoid structures. Its expression is regulated by inflammatory cytokine signaling and intersects with endothelial activation programs that coordinate leukocyte trafficking and local immune surveillance. Dysregulated MAdCAM-1 activity has been associated with chronic mucosal inflammation and immune-mediated pathology, making it a useful marker and mechanistic node for studying inflammation-driven tissue remodeling.

MAdCAM-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MADCAM1 expression without altering the underlying DNA sequence.

MAdCAM-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MADCAM1 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 MADCAM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MAdCAM-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MADCAM1 locus and enabling the study of MAdCAM-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MAdCAM-1 pathway restoration in tumor cells with silenced or reduced MADCAM1 expression.

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