NCAM/CD56 CRISPR Activation Plasmid (h): sc-400302-ACT

NCAM1 encodes the neural cell adhesion molecule (NCAM/CD56), an immunoglobulin superfamily glycoprotein that mediates homophilic and heterophilic cell–cell interactions and regulates adhesion-dependent signaling. NCAM/CD56 contributes to neurite outgrowth, synaptic plasticity, migration, and cytoskeletal remodeling through pathways involving Fyn/FAK, MAPK/ERK, and Rho-family GTPases, and it modulates cell-surface organization via polysialylation. In the immune system, NCAM/CD56 is a defining marker of natural killer cells and is linked to immune synapse formation and effector function. Dysregulated NCAM1 expression or glycosylation has been associated with neurodevelopmental and neurodegenerative phenotypes and is frequently studied in cancer biology where altered adhesion and invasion programs are implicated.

NCAM/CD56 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCAM1 expression without altering the underlying DNA sequence.

NCAM/CD56 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCAM1 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 NCAM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NCAM/CD56 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCAM1 locus and enabling the study of NCAM/CD56-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NCAM/CD56 pathway restoration in tumor cells with silenced or reduced NCAM1 expression.

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