Nicotinic Acetylcholine Receptor alpha 2/CHRNA2 CRISPR Activation Plasmid (h): sc-403376-ACT

CHRNA2 encodes the α2 subunit of neuronal nicotinic acetylcholine receptors (nAChRs), ligand-gated cation channels that assemble as pentamers to mediate rapid cholinergic neurotransmission. Upon acetylcholine or nicotine binding, CHRNA2-containing receptors drive Na⁺ and Ca²⁺ influx, influencing membrane excitability and activity-dependent signaling pathways that shape synaptic plasticity and circuit dynamics. In the central nervous system, altered nAChR subunit composition can modulate neurotransmitter release and downstream calcium-dependent transcriptional programs. Genetic and expression changes involving CHRNA2 have been investigated in the context of neuropsychiatric and neurodevelopmental phenotypes, where cholinergic signaling perturbations are implicated in disease-relevant neuronal network function.

Nicotinic Acetylcholine Receptor alpha 2/CHRNA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNA2 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor alpha 2/CHRNA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNA2 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 CHRNA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor alpha 2/CHRNA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNA2 locus and enabling the study of Nicotinic Acetylcholine Receptor alpha 2/CHRNA2-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 2/CHRNA2 pathway restoration in tumor cells with silenced or reduced CHRNA2 expression.

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