Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 CRISPR Activation Plasmid (h): sc-401702-ACT

CHRNA5 encodes the nicotinic acetylcholine receptor alpha 5 subunit, an accessory component of heteromeric ligand-gated cation channels that modulates receptor assembly, ion conductance, and desensitization kinetics in response to acetylcholine and nicotine. By shaping cholinergic signaling, CHRNA5 influences membrane depolarization, calcium-dependent signaling, and downstream neuronal excitability and synaptic plasticity pathways. Genetic variation and altered expression of CHRNA5 have been linked to susceptibility to nicotine dependence and smoking-related traits, and have been investigated in the context of lung epithelial biology and neuropsychiatric phenotypes. As a regulatory subunit, CHRNA5 can tune receptor responses in a cell-type-specific manner, making it relevant for mechanistic studies of cholinergic circuits and stimulus-dependent transcriptional programs.

Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNA5 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNA5 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 CHRNA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNA5 locus and enabling the study of Nicotinic Acetylcholine Receptor alpha 5/CHRNA5-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 5/CHRNA5 pathway restoration in tumor cells with silenced or reduced CHRNA5 expression.

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