ADAM23 CRISPR Activation Plasmid (h): sc-403703-ACT

ADAM23 encodes a disintegrin and metalloprotease domain-containing protein that functions primarily as a catalytically inactive cell-surface adhesion and signaling molecule. In human tissues, ADAM23 participates in cell–cell and cell–matrix interactions and has been linked to regulation of neuronal development, synaptic organization, and cell migration through interactions with extracellular ligands and integrin-associated pathways. Altered ADAM23 expression and epigenetic regulation have been reported in multiple disease contexts, including neurological disorders and cancer, where changes in adhesion signaling and invasive behavior are relevant. As a membrane-associated regulator of intercellular communication, ADAM23 provides a useful handle for dissecting how extracellular cues shape intracellular signaling programs.

ADAM23 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM23 expression without altering the underlying DNA sequence.

ADAM23 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM23 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 ADAM23 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAM23 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM23 locus and enabling the study of ADAM23-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAM23 pathway restoration in tumor cells with silenced or reduced ADAM23 expression.

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