KA2 CRISPR Activation Plasmid (h): sc-405272-ACT

GRIK5 encodes the kainate receptor subunit KA2, an obligate partner that assembles with other kainate receptor subunits to form functional ionotropic glutamate receptors at excitatory synapses. KA2-containing receptors contribute to fast synaptic transmission and activity-dependent plasticity by regulating cation flux and shaping neuronal excitability, linking GRIK5 to glutamatergic signaling networks and calcium-dependent downstream pathways. Through effects on synaptic integration and circuit-level excitation/inhibition balance, altered KA2 expression or receptor composition has been studied in the context of neurodevelopmental and neuropsychiatric phenotypes as well as seizure susceptibility. These properties make GRIK5 a useful node for interrogating synaptic receptor assembly, trafficking, and excitatory neurotransmission in human cellular models.

KA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GRIK5 expression without altering the underlying DNA sequence.

KA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GRIK5 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 GRIK5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GRIK5 locus and enabling the study of KA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KA2 pathway restoration in tumor cells with silenced or reduced GRIK5 expression.

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