Nicotinic Acetylcholine Receptor alpha 3/CHRNA3 CRISPR Activation Plasmid (h): sc-401582-ACT

CHRNA3 encodes the α3 subunit of neuronal nicotinic acetylcholine receptors, ligand-gated cation channels that mediate fast synaptic transmission and regulate membrane excitability through Na⁺ and Ca²⁺ influx. α3-containing nAChRs are prominent in autonomic ganglia and select neuronal circuits, where they influence neurotransmitter release, calcium-dependent signaling, and activity-dependent gene expression. Through cholinergic signaling, CHRNA3 contributes to pathways controlling neuronal differentiation, synaptic plasticity, and stimulus-secretion coupling. Genetic and expression studies have linked variation in the CHRNA3 locus and altered receptor activity to nicotine dependence and other neuropsychiatric phenotypes, supporting its relevance in addiction biology and neurocircuit function research.

Nicotinic Acetylcholine Receptor alpha 3/CHRNA3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNA3 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor alpha 3/CHRNA3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNA3 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 CHRNA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor alpha 3/CHRNA3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNA3 locus and enabling the study of Nicotinic Acetylcholine Receptor alpha 3/CHRNA3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nicotinic Acetylcholine Receptor alpha 3/CHRNA3 pathway restoration in tumor cells with silenced or reduced CHRNA3 expression.

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