DOR-1 CRISPR Activation Plasmid (h): sc-402195-ACT

OPRD1 encodes the human delta opioid receptor (DOR-1), a class A GPCR that couples primarily to Gi/Go proteins to regulate adenylyl cyclase activity, ion channel conductance, and MAPK signaling. DOR-1 modulates synaptic transmission and neuroimmune communication through downstream pathways such as cAMP/PKA and ERK1/2, shaping neuronal excitability and neurotransmitter release. Receptor activation influences processes including nociceptive signaling, stress responsivity, and reward-related circuitry, and altered OPRD1 expression or signaling has been associated with neuropsychiatric and pain-related phenotypes in biomedical research contexts. These features make OPRD1 a useful node for studying GPCR signaling dynamics, receptor trafficking/desensitization, and pathway bias in human cellular models.

DOR-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OPRD1 expression without altering the underlying DNA sequence.

DOR-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OPRD1 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 OPRD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DOR-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OPRD1 locus and enabling the study of DOR-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DOR-1 pathway restoration in tumor cells with silenced or reduced OPRD1 expression.

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