urocortin CRISPR Activation Plasmid (h): sc-403604-ACT

Human UCN encodes urocortin, a secreted neuropeptide of the corticotropin-releasing factor family that signals primarily through CRHR2 and CRHR1 to modulate cAMP/PKA-dependent transcriptional programs. Urocortin participates in neuroendocrine stress responses and coordinates cellular processes including secretion, metabolic adaptation, and inflammatory signaling across central and peripheral tissues. Through CRH receptor pathways, UCN influences vascular tone, gastrointestinal function, and cardiac physiology, linking its expression to studies of stress-related biology and cardiometabolic regulation. Altered urocortin signaling has been explored in contexts such as anxiety and depression-related phenotypes, heart failure, and inflammatory conditions, supporting mechanistic research into CRH axis dysregulation.

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

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

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