GABA_A Rδ CRISPR Activation Plasmid (h): sc-401954-ACT

GABRD encodes the δ subunit of the human GABA_A receptor, a ligand-gated chloride channel that mediates tonic inhibitory neurotransmission in the central nervous system. Incorporation of the δ subunit into extrasynaptic receptor complexes confers high sensitivity to ambient GABA and contributes to sustained chloride conductance that shapes neuronal excitability and network oscillations. Through regulation of inhibitory tone, GABA_A receptor δ signaling interfaces with synaptic plasticity, stress-responsive neurosteroid modulation, and excitation–inhibition balance. Altered GABRD expression or receptor composition has been linked in research to neuropsychiatric and neurological phenotypes involving disrupted inhibitory signaling, including seizure susceptibility and mood-related endophenotypes.

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

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

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