CASK CRISPR Activation Plasmid (h): sc-401742-ACT

CASK encodes a membrane-associated guanylate kinase (MAGUK) scaffold that organizes multiprotein complexes at synapses and cell–cell junctions, coupling adhesion molecules and ion channels to signaling and cytoskeletal regulators. In human neurons, CASK participates in synaptic vesicle trafficking, neurotransmission, and activity-dependent gene regulation, with roles linked to calcium/calmodulin signaling and junctional polarity programs. Perturbation of CASK dosage or function has been associated with neurodevelopmental phenotypes, including intellectual disability, epilepsy, and microcephaly, supporting its utility as a node for studying synapse assembly and neuronal maturation. These attributes make CASK a relevant target for dissecting pathways governing neuronal connectivity, excitability, and scaffold-dependent signaling networks.

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

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

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