CCS CRISPR Activation Plasmid (h): sc-402130-ACT

Human CCS (copper chaperone for superoxide dismutase) is a cytosolic metallochaperone that delivers copper to SOD1 and promotes its maturation, supporting antioxidant defense by enabling efficient dismutation of superoxide radicals. By regulating SOD1 metalation and stability, CCS contributes to redox homeostasis, mitochondrial function, and cellular resilience to oxidative stress across diverse cell types. Perturbations in copper handling and SOD1/CCS axis activity are relevant to neurodegeneration research and broader studies of oxidative stress–linked pathology, including inflammation and metabolic dysfunction. CCS expression and function are therefore commonly investigated in models examining copper-dependent signaling, proteostasis, and stress-response pathways.

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

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

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