ADAM9 CRISPR Activation Plasmid (h): sc-403665-ACT

ADAM9 encodes a transmembrane metalloprotease-disintegrin that regulates ectodomain shedding of membrane proteins and remodeling of the pericellular environment. Through proteolytic processing of growth factor precursors and adhesion molecules, ADAM9 influences cell–cell and cell–matrix interactions, receptor signaling, and proteostasis, interfacing with pathways linked to EGFR/ERBB signaling, integrin-mediated adhesion, and migratory programs. Dysregulated ADAM9 activity and expression have been associated with invasive phenotypes, inflammatory microenvironments, and altered tissue remodeling, making it a useful target for mechanistic studies of tumor biology and chronic disease-related remodeling processes.

ADAM9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM9 expression without altering the underlying DNA sequence.

ADAM9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM9 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 ADAM9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM9 locus and enabling the study of ADAM9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM9 pathway restoration in tumor cells with silenced or reduced ADAM9 expression.

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