GlyR α4 CRISPR Activation Plasmid (h): sc-416048-ACT

GLRA4 encodes the glycine receptor alpha 4 (GlyR α4), a ligand-gated chloride channel subunit that contributes to fast inhibitory neurotransmission in the central nervous system by mediating glycine-dependent hyperpolarization. GlyR complexes regulate neuronal excitability and network synchronization through chloride conductance and crosstalk with other inhibitory pathways, including GABAergic signaling. Although GLRA4 is less well characterized than other GLRA family members, glycinergic receptor biology is broadly linked to motor control, startle circuitry, and synaptic inhibitory balance, processes frequently implicated in neurodevelopmental and neuropsychiatric phenotypes. Studying GLRA4 supports mechanistic work on receptor assembly, synapse function, and inhibitory signaling alterations relevant to nervous system disorders without implying clinical outcomes.

GlyR α4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLRA4 expression without altering the underlying DNA sequence.

GlyR α4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLRA4 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 GLRA4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GlyR α4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLRA4 locus and enabling the study of GlyR α4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GlyR α4 pathway restoration in tumor cells with silenced or reduced GLRA4 expression.

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