NCAM2 CRISPR Activation Plasmid (h): sc-404803-ACT

NCAM2 (neural cell adhesion molecule 2) encodes a membrane-associated immunoglobulin superfamily adhesion protein enriched in the nervous system that supports neurite outgrowth, axon guidance, and synapse assembly. Through homophilic and heterophilic interactions at the cell surface, NCAM2 contributes to cell–cell recognition and stabilization of neuronal contacts, influencing cytoskeletal remodeling and activity-dependent connectivity. Altered NCAM2 expression or regulation has been implicated in neurodevelopmental and neuropsychiatric phenotypes, consistent with its role in shaping synaptic networks and neuronal circuit formation. As a marker and modulator of neuronal differentiation and plasticity, NCAM2 is broadly relevant for studies of adhesion-dependent signaling and synaptic organization.

NCAM2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCAM2 expression without altering the underlying DNA sequence.

NCAM2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCAM2 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 NCAM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NCAM2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCAM2 locus and enabling the study of NCAM2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NCAM2 pathway restoration in tumor cells with silenced or reduced NCAM2 expression.

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