Tyrosine Hydroxylase CRISPR Activation Plasmid (h): sc-400273-ACT

TH encodes tyrosine hydroxylase, the rate-limiting enzyme in catecholamine biosynthesis that converts L-tyrosine to L-DOPA, thereby controlling downstream dopamine, norepinephrine, and epinephrine production. Its activity integrates with tetrahydrobiopterin (BH4) cofactor metabolism and is regulated by phosphorylation-dependent signaling pathways that tune neurotransmitter output in response to neuronal activity and stress cues. TH expression and enzymatic control are central to dopaminergic and noradrenergic neuron identity, synaptic transmission, and neuroendocrine function. Dysregulation of TH and catecholamine homeostasis is implicated in neurological and neuropsychiatric disease biology, including disorders affecting dopaminergic circuitry and autonomic regulation.

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

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

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