ADAM12 CRISPR Activation Plasmid (h): sc-402583-ACT

ADAM12 encodes a membrane-anchored metalloprotease-disintegrin (ADAM12) that regulates cell–cell and cell–matrix interactions through ectodomain shedding of surface proteins and modulation of adhesion signaling. It participates in processes including extracellular matrix remodeling, myogenesis, and growth factor signaling crosstalk, influencing pathways such as EGFR/ERBB and TGF-β network activity via proteolytic processing of membrane-associated substrates. ADAM12 expression and activity have been linked to altered cellular migration, invasion, and differentiation programs in contexts of fibrosis and cancer biology. These properties make ADAM12 a useful target for studying protease-dependent signaling, stromal remodeling, and tissue development mechanisms in human cell models.

ADAM12 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM12 expression without altering the underlying DNA sequence.

ADAM12 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM12 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 ADAM12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM12 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM12 locus and enabling the study of ADAM12-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM12 pathway restoration in tumor cells with silenced or reduced ADAM12 expression.

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