D5DR CRISPR Activation Plasmid (h): sc-404047-ACT

DRD5 encodes the human dopamine receptor D5 (D5DR), a G protein-coupled receptor that primarily couples to Gs/olf to stimulate adenylyl cyclase, elevate cAMP, and activate PKA-dependent signaling. D5DR modulates neuronal excitability and synaptic plasticity through regulation of ion channels and phosphorylation networks, and it intersects with MAPK/ERK and other second-messenger pathways downstream of dopaminergic neurotransmission. DRD5 expression and signaling are studied in the context of dopaminergic circuit function, neurobehavioral phenotypes, and receptor cross-talk that shapes responses to catecholamine signaling. Dysregulated dopamine receptor signaling, including altered D5DR activity, is relevant to research on neuropsychiatric and neurodevelopmental disorders and on mechanisms of receptor desensitization and trafficking.

D5DR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DRD5 expression without altering the underlying DNA sequence.

D5DR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DRD5 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 DRD5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous D5DR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DRD5 locus and enabling the study of D5DR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of D5DR pathway restoration in tumor cells with silenced or reduced DRD5 expression.

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