TPH2 CRISPR Activation Plasmid (h): sc-402296-ACT

TPH2 (tryptophan hydroxylase 2) encodes the neuronal, rate-limiting enzyme for serotonin (5‑hydroxytryptamine) biosynthesis, catalyzing the hydroxylation of L‑tryptophan to 5‑hydroxy‑L‑tryptophan in serotonergic neurons. As a key determinant of serotonin availability, TPH2 influences neurotransmission programs that modulate mood, stress responsivity, sleep–wake regulation, nociception, and autonomic function. Altered TPH2 expression or genetic variation has been investigated in the context of neuropsychiatric and neurodevelopmental phenotypes where serotonergic signaling is implicated, including affective and anxiety-related traits. In cellular models, TPH2 activity intersects with pathways controlling monoamine metabolism, synaptic plasticity, and activity-dependent gene regulation.

TPH2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TPH2 expression without altering the underlying DNA sequence.

TPH2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TPH2 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 TPH2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TPH2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TPH2 locus and enabling the study of TPH2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TPH2 pathway restoration in tumor cells with silenced or reduced TPH2 expression.

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