SR-2C CRISPR Activation Plasmid (h): sc-400886-ACT

HTR2C encodes the human 5-hydroxytryptamine receptor 2C (SR-2C), a G protein–coupled receptor that primarily signals through Gq/11 to activate phospholipase C, increase intracellular Ca2+ mobilization, and engage PKC-dependent phosphorylation cascades. SR-2C also influences MAPK/ERK signaling and regulates neurotransmitter release through modulation of neuronal excitability and synaptic transmission in serotonergic circuits. Through these pathways, HTR2C contributes to central control of appetite, mood, and endocrine/autonomic outputs, and its dysregulation has been linked to neuropsychiatric and metabolic phenotypes in experimental and genetic studies. These features make HTR2C a useful target for investigating GPCR signaling dynamics, receptor editing/isoform effects, and pathway crosstalk in neuronal and neuroendocrine cell models.

SR-2C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HTR2C expression without altering the underlying DNA sequence.

SR-2C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HTR2C 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 HTR2C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SR-2C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HTR2C locus and enabling the study of SR-2C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SR-2C pathway restoration in tumor cells with silenced or reduced HTR2C expression.

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