GluR-5 CRISPR Activation Plasmid (h): sc-402475-ACT

GRIK1 encodes the kainate-type ionotropic glutamate receptor subunit GluR-5 (GluK1), a ligand-gated cation channel that contributes to fast excitatory neurotransmission in the central nervous system. GluR-5-containing receptors modulate synaptic strength and neuronal excitability through sodium and calcium flux, shaping network activity and plasticity in pathways involved in learning, sensory processing, and pain signaling. Receptor activity interfaces with calcium-dependent signaling cascades and activity-regulated transcriptional programs that influence synapse development and remodeling. Dysregulated kainate receptor signaling and GRIK1 expression have been associated with neuropsychiatric and neurological phenotypes, supporting its use as a mechanistic target for studying excitatory/inhibitory balance and circuit dysfunction.

GluR-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRIK1 expression without altering the underlying DNA sequence.

GluR-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRIK1 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 GRIK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GluR-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRIK1 locus and enabling the study of GluR-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GluR-5 pathway restoration in tumor cells with silenced or reduced GRIK1 expression.

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