GlyR α2 CRISPR Activation Plasmid (h): sc-402179-ACT

GLRA2 encodes the human glycine receptor alpha-2 subunit (GlyR α2), a ligand-gated chloride channel that mediates fast inhibitory neurotransmission in the central nervous system. Upon glycine binding, GlyR α2 promotes chloride influx and membrane hyperpolarization, shaping neuronal excitability and synaptic signaling during development and in mature circuits. GLRA2-linked alterations have been associated with neurodevelopmental phenotypes and seizure-related disorders, highlighting the importance of glycinergic signaling in maintaining inhibitory tone. This receptor interfaces functionally with broader neurotransmitter networks and ion homeostasis processes that regulate circuit maturation and excitatory–inhibitory balance.

GlyR α2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLRA2 expression without altering the underlying DNA sequence.

GlyR α2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLRA2 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 GLRA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GlyR α2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLRA2 locus and enabling the study of GlyR α2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GlyR α2 pathway restoration in tumor cells with silenced or reduced GLRA2 expression.

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