ceruloplasmin CRISPR Activation Plasmid (h): sc-401798-ACT

Human CP encodes ceruloplasmin, a secreted multicopper ferroxidase that catalyzes Fe2+ to Fe3+ conversion to support transferrin loading and systemic iron trafficking. Through its role in ferroxidase activity and copper binding, ceruloplasmin helps maintain redox balance and limits iron-driven oxidative stress, linking CP to metal homeostasis pathways in hepatocytes, macrophages, and the nervous system. Altered CP expression or activity has been associated with dysregulated iron accumulation, inflammatory states, and neurodegenerative phenotypes where oxidative damage and impaired iron export contribute to pathology. These features make CP a useful node for studying ferroptosis susceptibility, iron-dependent signaling, and secreted protein contributions to extracellular redox biology.

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

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

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