NCAM-L1 CRISPR Activation Plasmid (h): sc-401012-ACT

L1CAM encodes NCAM-L1 (L1), a neuronal cell adhesion molecule that mediates homophilic and heterophilic interactions to regulate neurite outgrowth, axon guidance, neuronal migration, and synaptic plasticity. Through coupling to cytoskeletal adaptors and signaling nodes, NCAM-L1 influences pathways controlling adhesion dynamics, growth cone motility, and MAPK/ERK- and PI3K-linked processes that shape neural circuit development. In the nervous system, altered L1CAM expression or function is associated with neurodevelopmental phenotypes and has also been reported in contexts of tumor cell motility and invasive behavior, supporting its value for mechanistic studies of adhesion-dependent signaling. These properties make L1CAM a useful target for dissecting cell–cell interaction programs and their downstream transcriptional and morphological outputs.

NCAM-L1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous L1CAM expression without altering the underlying DNA sequence.

NCAM-L1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the L1CAM 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 L1CAM transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NCAM-L1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native L1CAM locus and enabling the study of NCAM-L1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NCAM-L1 pathway restoration in tumor cells with silenced or reduced L1CAM expression.

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