NMDAε2 CRISPR Activation Plasmid (h): sc-400654-ACT

GRIN2B encodes the human NMDA receptor subunit NMDAε2 (GluN2B), a glutamate-gated ion channel component that regulates Ca²⁺ influx during excitatory neurotransmission. NMDAε2-containing receptors shape synaptic plasticity, activity-dependent gene expression, and neuronal development through coupling to CaMK/CREB signaling, MAPK pathways, and downstream transcriptional programs. GRIN2B function is tightly linked to excitatory–inhibitory balance, dendritic spine maturation, and circuit refinement in the central nervous system. Altered GRIN2B expression or NMDA receptor signaling has been associated with neurodevelopmental and neuropsychiatric disease mechanisms, including cognitive dysfunction, epilepsy-related phenotypes, and synaptopathies.

NMDAε2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRIN2B expression without altering the underlying DNA sequence.

NMDAε2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRIN2B 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 GRIN2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NMDAε2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRIN2B locus and enabling the study of NMDAε2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NMDAε2 pathway restoration in tumor cells with silenced or reduced GRIN2B expression.

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