Nicotinic Acetylcholine Receptor alpha 7/CHRNA7 CRISPR Activation Plasmid (h): sc-416614-ACT

CHRNA7 encodes the α7 subunit of the nicotinic acetylcholine receptor, a homopentameric ligand-gated cation channel that mediates rapid Ca²⁺ influx in response to cholinergic signaling. In neurons and immune cells, α7 nAChR activity influences membrane excitability and calcium-dependent signaling cascades that shape synaptic plasticity, neurotransmitter release, and transcriptional programs linked to the cholinergic anti-inflammatory pathway. CHRNA7-regulated calcium signaling interfaces with MAPK/ERK, JAK/STAT, and NF-κB-associated processes, impacting cytokine production and cellular stress responses. Genetic variation and altered expression of CHRNA7 have been associated with neuropsychiatric and neurodevelopmental phenotypes, and dysregulated cholinergic signaling is also studied in contexts of inflammation and neurodegeneration.

Nicotinic Acetylcholine Receptor alpha 7/CHRNA7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNA7 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor alpha 7/CHRNA7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNA7 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 CHRNA7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor alpha 7/CHRNA7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNA7 locus and enabling the study of Nicotinic Acetylcholine Receptor alpha 7/CHRNA7-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 7/CHRNA7 pathway restoration in tumor cells with silenced or reduced CHRNA7 expression.

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