SR-1B CRISPR Activation Plasmid (h): sc-410541-ACT

HTR1B encodes the human 5-hydroxytryptamine receptor 1B (SR-1B), a Gi/o-coupled GPCR that modulates neurotransmitter release and neuronal excitability by inhibiting adenylyl cyclase, reducing cAMP signaling, and regulating ion channel activity. SR-1B signaling influences synaptic transmission within serotonergic circuits and integrates with pathways controlling dopamine and GABA release, shaping reward processing, impulse control, and stress responses. Altered HTR1B expression or signaling has been associated with neuropsychiatric phenotypes and behavioral endophenotypes, supporting its use in mechanistic studies of serotonergic pathway regulation. As a membrane receptor with context-dependent expression, SR-1B provides a tractable node for dissecting GPCR-driven transcriptional programs and downstream second-messenger dynamics in human cellular models.

SR-1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HTR1B expression without altering the underlying DNA sequence.

SR-1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HTR1B 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 HTR1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SR-1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HTR1B locus and enabling the study of SR-1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SR-1B pathway restoration in tumor cells with silenced or reduced HTR1B expression.

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