CPE CRISPR Activation Plasmid (h): sc-404429-ACT

Carboxypeptidase E (CPE) is a zinc-dependent exopeptidase that processes C-terminal basic residues from peptide hormone and neuropeptide precursors, supporting the maturation of bioactive secretory peptides. In neuroendocrine and endocrine cells, CPE functions within the regulated secretory pathway and is linked to vesicle sorting and dense-core granule biogenesis, influencing peptide trafficking and stimulus-coupled secretion. By shaping the availability of signaling peptides, CPE impacts pathways governing metabolism, neuronal communication, and stress responses. Altered CPE expression or activity has been associated with neuroendocrine dysfunction and metabolic phenotypes, making it a relevant target for studies of peptide processing networks and secretory pathway regulation.

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

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

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