Nicotinic Acetylcholine Receptor alpha 9/CHRNA9 CRISPR Activation Plasmid (h): sc-402753-ACT

CHRNA9 encodes the nicotinic acetylcholine receptor alpha 9 subunit, a ligand-gated ion channel component that forms functional receptors (often with CHRNA10) to mediate cholinergic signaling and rapid cation flux across the plasma membrane. Through acetylcholine-dependent channel gating, CHRNA9 influences membrane excitability, calcium-dependent signaling cascades, and downstream transcriptional programs that shape cellular communication. In non-neuronal contexts, nicotinic receptor signaling has been linked to regulation of proliferation, differentiation, and inflammatory signaling, positioning CHRNA9 as a useful node for studying stimulus-coupled ion channel pathways. Altered CHRNA9 expression or signaling has been associated with disease-relevant phenotypes reported in sensory biology and cancer-related cell behaviors, supporting its investigation in mechanistic models.

Nicotinic Acetylcholine Receptor alpha 9/CHRNA9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNA9 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor alpha 9/CHRNA9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNA9 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 CHRNA9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor alpha 9/CHRNA9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNA9 locus and enabling the study of Nicotinic Acetylcholine Receptor alpha 9/CHRNA9-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 9/CHRNA9 pathway restoration in tumor cells with silenced or reduced CHRNA9 expression.

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