Nicotinic Acetylcholine Receptor beta 1/CHRNB1 CRISPR Activation Plasmid (h): sc-404205-ACT

CHRNB1 encodes the β1 subunit of the muscle-type nicotinic acetylcholine receptor (nAChR), a ligand-gated ion channel that assembles with α, δ, and ε/γ subunits to mediate fast synaptic transmission at the neuromuscular junction. Upon acetylcholine binding, the receptor opens to permit cation flux, linking cholinergic signaling to membrane depolarization, excitation–contraction coupling, and activity-dependent remodeling of postsynaptic architecture. CHRNB1 function is integrated with agrin–LRP4–MuSK signaling and acetylcholinesterase-regulated synaptic clearance that together maintain endplate stability and muscle excitability. Genetic or autoimmune perturbations affecting nAChR integrity are associated with congenital myasthenic syndromes and myasthenia gravis–related mechanisms, making CHRNB1 a key target for studying neuromuscular transmission defects and receptor assembly biology.

Nicotinic Acetylcholine Receptor beta 1/CHRNB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHRNB1 expression without altering the underlying DNA sequence.

Nicotinic Acetylcholine Receptor beta 1/CHRNB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHRNB1 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 CHRNB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nicotinic Acetylcholine Receptor beta 1/CHRNB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHRNB1 locus and enabling the study of Nicotinic Acetylcholine Receptor beta 1/CHRNB1-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 beta 1/CHRNB1 pathway restoration in tumor cells with silenced or reduced CHRNB1 expression.

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