RIC-3 CRISPR Activation Plasmid (h): sc-405366-ACT

Human RIC3 encodes RIC-3, an endoplasmic reticulum resident chaperone that promotes folding, assembly, and trafficking of select ligand-gated ion channels, most notably nicotinic acetylcholine receptors. By controlling receptor maturation and surface expression, RIC-3 influences cholinergic synaptic transmission, neuronal excitability, and downstream calcium-dependent signaling programs. Altered regulation of nicotinic receptor biogenesis and cholinergic network function has been linked to neurological and neuropsychiatric phenotypes, making RIC3 a useful node for studying receptor proteostasis and synaptic signaling. RIC-3 is also leveraged as a mechanistic handle to dissect ER quality control, channel complex stoichiometry, and stimulus-dependent changes in receptor availability.

RIC-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RIC3 expression without altering the underlying DNA sequence.

RIC-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RIC3 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 RIC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RIC-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RIC3 locus and enabling the study of RIC-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RIC-3 pathway restoration in tumor cells with silenced or reduced RIC3 expression.

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