secretin CRISPR Activation Plasmid (m): sc-422831-ACT

Mouse Sct encodes secretin, a peptide hormone primarily produced by enteroendocrine S cells in the duodenum that coordinates digestive and metabolic physiology. Secretin signals through the secretin receptor (a class B GPCR) to activate adenylate cyclase and cAMP/PKA pathways, regulating pancreatic bicarbonate secretion, bile flow, and gastric acid inhibition, and contributing to water and electrolyte homeostasis. In the nervous system, secretin has been implicated in neuromodulatory processes, including effects on synaptic activity and hypothalamic–pituitary signaling. Dysregulated secretin/secretin receptor signaling is studied in contexts such as gastrointestinal dysfunction, pancreatic exocrine insufficiency, cholestatic processes, and neuroendocrine pathway perturbations.

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

secretin CRISPR Activation Plasmid (m) 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.