secretin CRISPR Activation Plasmid (h): sc-405534-ACT

Human SCT encodes secretin, a peptide hormone classically produced by duodenal S cells that coordinates gastrointestinal physiology by stimulating pancreatic and biliary bicarbonate secretion and modulating gastric acid output. Secretin signals primarily through the secretin receptor (SCTR), a G protein–coupled receptor that activates adenylate cyclase and cAMP/PKA-dependent transcriptional programs in target epithelia. Beyond digestive functions, secretin-responsive pathways influence fluid and electrolyte balance and can shape neuroendocrine signaling in select tissues. Dysregulated secretin–SCTR axis activity has been investigated in the context of pancreatic and biliary disorders and broader endocrine-metabolic phenotypes, supporting mechanistic studies of hormone-regulated epithelial transport and secretion.

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

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

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