OR2A1 CRISPR Activation Plasmid (h): sc-417340-ACT

Human OR2A1 encodes an olfactory receptor in the class A GPCR family that couples to olfactory G protein signaling to regulate odorant-evoked sensory transduction. Upon ligand binding, OR2A1 typically engages adenylate cyclase–cAMP pathways to modulate cyclic nucleotide–gated ion channel activity and downstream neuronal excitability programs. Although best characterized in the olfactory epithelium, olfactory receptor transcripts have been reported in diverse peripheral tissues, motivating studies of ectopic GPCR signaling and context-specific transcriptional regulation. Altered expression patterns across tissues and disease states have made OR2A1 a useful marker gene for investigating sensory GPCR gene regulation and chemosensory-related phenotypes in cellular models.

OR2A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OR2A1 expression without altering the underlying DNA sequence.

OR2A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OR2A1 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 OR2A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OR2A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OR2A1 locus and enabling the study of OR2A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OR2A1 pathway restoration in tumor cells with silenced or reduced OR2A1 expression.

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