ADAM15 CRISPR Activation Plasmid (h): sc-405627-ACT

Human ADAM15 encodes a membrane-anchored metalloprotease-disintegrin (ADAM) family protein that integrates proteolytic shedding with cell–cell and cell–matrix interactions. ADAM15 contributes to ectodomain cleavage of select surface proteins and modulates integrin-dependent adhesion, influencing cytoskeletal remodeling, migration, and inflammatory signaling programs. Through these activities, ADAM15 can impact pathways linked to extracellular matrix turnover, angiogenic responses, and receptor signaling dynamics at the cell surface. Dysregulated ADAM15 expression has been associated with tumor progression and invasive phenotypes, as well as vascular and inflammatory pathobiology, making it relevant for mechanistic studies of microenvironment-driven signaling.

ADAM15 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM15 expression without altering the underlying DNA sequence.

ADAM15 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM15 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 ADAM15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM15 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM15 locus and enabling the study of ADAM15-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM15 pathway restoration in tumor cells with silenced or reduced ADAM15 expression.

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