GABA_A Rα3 CRISPR Activation Plasmid (h): sc-403044-ACT

GABRA3 encodes the human γ-aminobutyric acid type A receptor alpha 3 subunit (GABAA Rα3), a ligand-gated chloride channel component that mediates fast inhibitory neurotransmission. Incorporation of the α3 subunit into pentameric GABAA receptors shapes channel gating, pharmacology, and synaptic versus extrasynaptic inhibitory tone, influencing neuronal excitability and network oscillations. GABAA Rα3 participates in GABAergic synapse function and broader neurodevelopmental processes including neurite outgrowth and circuit maturation through activity-dependent signaling. Altered GABRA3 expression or inhibitory signaling imbalance has been associated in research with neurological phenotypes and disease-relevant states where disrupted excitation–inhibition balance contributes to pathophysiology.

GABAA Rα3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GABRA3 expression without altering the underlying DNA sequence.

GABAA Rα3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GABRA3 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 GABRA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GABAA Rα3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GABRA3 locus and enabling the study of GABAA Rα3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GABAA Rα3 pathway restoration in tumor cells with silenced or reduced GABRA3 expression.

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