DBH CRISPR Activation Plasmid (h): sc-402441-ACT

Dopamine beta-hydroxylase (DBH) encodes a copper-dependent monooxygenase that catalyzes the conversion of dopamine to norepinephrine, a key step in catecholamine biosynthesis within sympathetic neurons and adrenal chromaffin cells. DBH activity supports vesicular neurotransmitter metabolism and integrates with tyrosine hydroxylase–driven pathways that regulate adrenergic signaling, neuroendocrine stress responses, and cardiovascular homeostasis. Altered DBH expression or enzymatic function has been linked to dysregulated norepinephrine levels and is studied in the context of neuropsychiatric traits and autonomic dysfunction. As a molecular readout of catecholaminergic state, DBH is widely used to probe noradrenergic lineage identity, neuronal maturation, and stimulus-dependent neurotransmitter remodeling.

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

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

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