Neurogranin CRISPR Activation Plasmid (h): sc-404119-ACT

Human NRGN encodes neurogranin, a postsynaptic calmodulin-binding protein enriched in dendritic spines that tunes Ca2+/calmodulin availability and supports activity-dependent synaptic signaling. By modulating CaMKII-dependent phosphorylation dynamics and downstream plasticity programs, neurogranin contributes to long-term potentiation, spine remodeling, and neuronal circuit maturation. NRGN expression and neurogranin phosphorylation state are linked to glutamatergic transmission and broader pathways governing learning and memory, including CREB-regulated transcriptional responses. Dysregulation of neurogranin biology has been associated with neuropsychiatric and neurodegenerative disease-relevant phenotypes, making it a useful node for mechanistic studies of synaptic dysfunction.

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

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

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