Arc CRISPR Activation Plasmid (h): sc-400231-ACT

ARC encodes activity-regulated cytoskeleton-associated protein (Arc), an immediate-early gene product that couples neuronal activity to synaptic remodeling. Arc regulates AMPA receptor endocytosis, actin cytoskeleton dynamics, and local translation at dendritic spines, thereby shaping synaptic plasticity, learning, and memory. Through these functions it intersects with activity-dependent signaling pathways and homeostatic synaptic scaling programs that tune excitatory neurotransmission. Dysregulated ARC expression or Arc-dependent trafficking has been implicated in neuropsychiatric and neurodegenerative research contexts, including studies of cognitive dysfunction, seizure susceptibility, and altered synapse density.

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

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

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