SR-7 CRISPR Activation Plasmid (h): sc-404174-ACT

HTR7 encodes the human 5-hydroxytryptamine receptor 7 (SR-7), a G protein–coupled receptor that primarily couples to Gαs to elevate intracellular cAMP and engage PKA-dependent signaling. SR-7 modulates neuronal excitability and synaptic plasticity and can influence downstream transcriptional programs through cAMP/CREB as well as cross-talk with ERK/MAPK pathways. In addition to roles in central nervous system circuitry, HTR7 signaling has been linked to regulation of circadian rhythms, mood-associated behaviors, and neuroinflammatory responses, supporting its use as a mechanistic node in neuropsychiatric and neurobiology research. Altered serotonergic GPCR signaling involving HTR7 has also been investigated in contexts of sleep and affective phenotypes, making it relevant for pathway-focused studies of neurotransmitter signaling and cellular homeostasis.

SR-7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HTR7 expression without altering the underlying DNA sequence.

SR-7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HTR7 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 HTR7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SR-7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HTR7 locus and enabling the study of SR-7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SR-7 pathway restoration in tumor cells with silenced or reduced HTR7 expression.

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