ADAM28 CRISPR Activation Plasmid (h): sc-404031-ACT

ADAM28 encodes a membrane-anchored metalloprotease of the ADAM (a disintegrin and metalloprotease) family that mediates ectodomain shedding and cell–cell or cell–matrix interactions through its disintegrin and protease domains. By regulating the availability of surface receptors, adhesion molecules, and soluble mediators, ADAM28 influences proteolysis-dependent signaling, leukocyte trafficking, and tissue remodeling. ADAM28 expression has been studied in contexts involving inflammatory cell recruitment and remodeling of the tumor microenvironment, where altered protease activity can shift adhesion dynamics and cytokine responsiveness. As a surface-associated protease, ADAM28 is frequently interrogated in pathways linked to extracellular matrix regulation, immune signaling, and migratory phenotypes.

ADAM28 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM28 expression without altering the underlying DNA sequence.

ADAM28 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM28 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 ADAM28 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM28 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM28 locus and enabling the study of ADAM28-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM28 pathway restoration in tumor cells with silenced or reduced ADAM28 expression.

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