ADAM8 CRISPR Activation Plasmid (h): sc-407606-ACT

ADAM8 encodes a membrane-anchored metalloprotease-disintegrin that mediates ectodomain shedding and cell–cell or cell–matrix interactions, linking proteolysis to adhesion-dependent signaling. By processing select surface receptors, cytokines, and extracellular matrix components, ADAM8 can influence leukocyte trafficking, inflammatory signaling, and remodeling programs that shape tissue microenvironments. Its activity intersects with pathways governing migration and invasion, including integrin-related signaling and protease networks that regulate extracellular matrix turnover. Dysregulated ADAM8 expression has been associated with inflammatory conditions and multiple cancer contexts, making it a useful node for studying protease-driven communication between immune and stromal compartments.

ADAM8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM8 expression without altering the underlying DNA sequence.

ADAM8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM8 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 ADAM8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM8 locus and enabling the study of ADAM8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM8 pathway restoration in tumor cells with silenced or reduced ADAM8 expression.

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