CSN8 CRISPR Activation Plasmid (h): sc-409800-ACT

COPS8 encodes CSN8, a core subunit of the COP9 signalosome that regulates cullin-RING ubiquitin ligase activity through deneddylation, thereby shaping ubiquitin-dependent protein turnover. By controlling the stability of key cell-cycle regulators, DNA damage response factors, and stress signaling intermediates, CSN8 helps coordinate proteostasis, proliferation, and adaptation to environmental cues. CSN8 function intersects with pathways linked to apoptosis, autophagy, and inflammatory signaling, reflecting the central role of COP9 signalosome activity in cellular homeostasis. Dysregulated CSN8-associated ubiquitin signaling has been implicated in disease-relevant mechanisms such as aberrant cell-cycle progression, altered proteasome function, and stress pathway imbalance in multiple tissue contexts.

CSN8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COPS8 expression without altering the underlying DNA sequence.

CSN8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COPS8 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 COPS8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CSN8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COPS8 locus and enabling the study of CSN8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CSN8 pathway restoration in tumor cells with silenced or reduced COPS8 expression.

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