R7BP CRISPR Activation Plasmid (h): sc-415971-ACT

RGS7BP encodes R7BP, a membrane-anchoring auxiliary subunit that binds R7 family RGS proteins such as RGS7/RGS9 and modulates their localization and activity in G protein–coupled receptor (GPCR) signaling. By promoting membrane targeting and complex stability, R7BP influences Gi/o-mediated signal termination and downstream second-messenger dynamics, including cAMP and ion channel regulation in excitable cells. This regulatory axis is implicated in neuronal signal integration and synaptic plasticity, and altered RGS7BP-associated signaling has been connected to neuropsychiatric and neurodegenerative disease biology through dysregulated GPCR pathway control. RGS7BP is therefore used as a molecular entry point to interrogate GPCR signal kinetics, membrane trafficking, and neuronal network responsiveness.

R7BP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RGS7BP expression without altering the underlying DNA sequence.

R7BP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RGS7BP 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 RGS7BP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous R7BP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RGS7BP locus and enabling the study of R7BP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of R7BP pathway restoration in tumor cells with silenced or reduced RGS7BP expression.

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