secretin receptor CRISPR Activation Plasmid (h): sc-403329-ACT

SCTR encodes the human secretin receptor, a class B G protein–coupled receptor that binds the gastrointestinal peptide hormone secretin to regulate epithelial fluid and bicarbonate secretion, pancreatic ductal function, and biliary physiology. Upon ligand engagement, SCTR primarily couples to Gs to stimulate adenylyl cyclase, elevate cAMP, and activate PKA-dependent transcriptional and ion-transport programs, with additional context-dependent signaling through calcium and MAPK pathways. Receptor activity contributes to coordinated digestive and metabolic homeostasis and is frequently studied in secretory epithelia and neuroendocrine contexts. Altered GPCR/cAMP signaling involving SCTR has been investigated in models of pancreatic and biliary dysfunction, gastrointestinal disorders, and hormone-responsive tumor biology as a mechanism shaping cellular secretion, proliferation, and differentiation.

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

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

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