ADAM10 CRISPR Activation Plasmid (h): sc-400883-ACT

ADAM10 encodes a membrane-anchored metalloprotease of the ADAM (a disintegrin and metalloprotease) family that functions as a major α-secretase responsible for ectodomain shedding of diverse cell-surface proteins. By processing substrates such as NOTCH receptors/ligands, cadherins, cytokine receptors, and amyloid precursor protein, ADAM10 influences cell fate decisions, adhesion, migration, and inflammatory signaling across multiple tissues. ADAM10-mediated proteolysis modulates Notch signaling dynamics and membrane receptor availability, shaping downstream transcriptional programs and intercellular communication. Dysregulated ADAM10 activity has been associated with pathological remodeling of signaling networks implicated in neurodegeneration, cancer biology, and immune dysregulation, making it a useful target for mechanistic studies of protease-dependent pathway control.

ADAM10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM10 expression without altering the underlying DNA sequence.

ADAM10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM10 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 ADAM10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM10 locus and enabling the study of ADAM10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM10 pathway restoration in tumor cells with silenced or reduced ADAM10 expression.

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