D4DR CRISPR Activation Plasmid (h): sc-402947-ACT

Human DRD4 encodes the dopamine receptor D4 (D4DR), a Gi/o-coupled GPCR that modulates adenylate cyclase activity, cAMP/PKA signaling, and downstream phosphorylation programs that shape neuronal excitability and synaptic transmission. D4DR also engages MAPK and β-arrestin–linked pathways, influencing receptor desensitization, trafficking, and longer-term transcriptional responses to dopaminergic tone. DRD4 expression and regulatory variation have been investigated in the context of frontostriatal circuit function and neuropsychiatric phenotypes, making it a useful molecular node for studying dopaminergic signaling dynamics. In cellular models, DRD4 perturbation can be used to probe pathway coupling, receptor pharmacology, and transcriptional responses to dopamine in relevant neural and heterologous expression systems.

D4DR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DRD4 expression without altering the underlying DNA sequence.

D4DR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DRD4 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 DRD4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous D4DR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DRD4 locus and enabling the study of D4DR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of D4DR pathway restoration in tumor cells with silenced or reduced DRD4 expression.

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