SR-2B CRISPR Activation Plasmid (h): sc-402574-ACT

Human HTR2B encodes the serotonin receptor 2B (SR-2B), a G protein–coupled receptor that primarily couples to Gq/11 to stimulate phospholipase C signaling, inositol phosphate turnover, intracellular Ca2+ mobilization, and PKC-dependent MAPK/ERK pathway activity. SR-2B modulates transcriptional programs linked to cell growth, extracellular matrix remodeling, and cytoskeletal dynamics, integrating serotonergic cues with tissue-specific signaling networks. Expression and signaling changes in HTR2B have been associated with cardiovascular remodeling and valvular biology, and altered receptor activity has been reported across neuropsychiatric and fibrotic contexts, supporting mechanistic studies of serotonin-driven pathophysiology. These features make SR-2B a useful target for investigating GPCR signal transduction, receptor-biased signaling, and downstream gene-regulatory responses in human cell models.

SR-2B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HTR2B expression without altering the underlying DNA sequence.

SR-2B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HTR2B 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 HTR2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SR-2B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HTR2B locus and enabling the study of SR-2B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SR-2B pathway restoration in tumor cells with silenced or reduced HTR2B expression.

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