SR-2B CRISPR Activation Plasmid (m): sc-420994-ACT

Mouse Htr2b encodes the serotonin receptor SR-2B (5-HT2B), a G protein–coupled receptor that primarily signals through Gq/11 to activate phospholipase C, inositol phosphate turnover, intracellular Ca²⁺ mobilization, and downstream PKC/ERK transcriptional programs. SR-2B modulates neuromodulatory and peripheral serotonergic signaling that influences neuronal excitability, vascular tone, and tissue remodeling responses. Altered Htr2b activity has been associated with dysregulated serotonin pathway outputs relevant to neurobehavioral phenotypes, cardiovascular and pulmonary remodeling processes, and inflammatory signaling contexts. As a membrane receptor node, SR-2B provides a tractable entry point for studying receptor-driven gene expression, second-messenger coupling, and pathway cross-talk with MAPK and calcium-dependent networks.

SR-2B CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Htr2b expression without altering the underlying DNA sequence.

SR-2B CRISPR Activation Plasmid (m) 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.