SR-1D CRISPR Activation Plasmid (h): sc-404213-ACT

HTR1D encodes the human 5-hydroxytryptamine receptor 1D (SR-1D), a Gi/o-coupled GPCR that binds serotonin to inhibit adenylyl cyclase, reduce cAMP signaling, and modulate neuronal excitability and neurotransmitter release. SR-1D participates in serotonergic synaptic transmission and downstream pathways including cAMP/PKA, MAPK/ERK, and ion channel regulation, with functional effects shaped by receptor trafficking and desensitization. Expression and signaling changes in HTR1D have been investigated in neurovascular and neuropsychiatric biology, including mechanisms relevant to headache pathophysiology and pain processing. As a presynaptic modulator, SR-1D provides a tractable node for dissecting GPCR signaling bias, circuit-level neurotransmission, and ligand-dependent receptor dynamics in human cell models.

SR-1D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HTR1D expression without altering the underlying DNA sequence.

SR-1D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HTR1D 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 HTR1D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SR-1D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HTR1D locus and enabling the study of SR-1D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SR-1D pathway restoration in tumor cells with silenced or reduced HTR1D expression.

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