CASPR CRISPR Activation Plasmid (h): sc-402760-ACT

CNTNAP1 encodes contactin-associated protein 1 (CASPR), a neurexin superfamily cell-adhesion molecule concentrated at paranodal junctions of myelinated axons. CASPR coordinates axon–glia interactions by organizing septate-like junctions and promoting proper localization of ion channels required for efficient saltatory conduction. Through its extracellular adhesion domains and intracellular scaffolding interactions, CASPR contributes to the assembly and maintenance of nodes of Ranvier architecture and overall neuronal circuit integrity. Disruption of CNTNAP1 function has been associated with severe neurodevelopmental phenotypes and peripheral neuropathies, making it a relevant target for studying myelination biology and axonal domain organization.

CASPR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CNTNAP1 expression without altering the underlying DNA sequence.

CASPR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CNTNAP1 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 CNTNAP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CASPR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CNTNAP1 locus and enabling the study of CASPR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CASPR pathway restoration in tumor cells with silenced or reduced CNTNAP1 expression.

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