GluR-δ2 CRISPR Activation Plasmid (h): sc-403400-ACT

GRID2 encodes the ionotropic glutamate receptor delta-2 (GluR-δ2), a cerebellum-enriched receptor-like protein that is essential for excitatory synapse organization and synaptic plasticity. Rather than functioning as a conventional ligand-gated channel, GluR-δ2 acts as a synaptic organizer at parallel fiber–Purkinje cell synapses, coordinating receptor signaling complexes and activity-dependent remodeling. It contributes to processes such as long-term depression, dendritic spine maturation, and cerebellar circuit development through glutamatergic signaling networks. Dysregulation of GRID2 has been linked to neurodevelopmental and neurodegenerative phenotypes characterized by cerebellar dysfunction, including ataxia-associated pathways.

GluR-δ2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRID2 expression without altering the underlying DNA sequence.

GluR-δ2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRID2 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 GRID2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GluR-δ2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRID2 locus and enabling the study of GluR-δ2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GluR-δ2 pathway restoration in tumor cells with silenced or reduced GRID2 expression.

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