CSN6 CRISPR Activation Plasmid (h): sc-404057-ACT

Human COPS6 encodes CSN6, a core subunit of the COP9 signalosome that regulates cullin-RING E3 ubiquitin ligases through control of cullin neddylation and deneddylation dynamics. By modulating CRL activity, CSN6 influences ubiquitin-dependent proteostasis, cell-cycle progression, DNA damage signaling, and stress-responsive transcriptional programs. CSN6 has been linked to altered stability of key regulatory proteins, connecting COP9 signalosome function to oncogenic signaling networks and other disorders characterized by disrupted protein turnover. These properties make COPS6 a relevant target for studying ubiquitin–proteasome system crosstalk with signal transduction and chromatin-associated regulation.

CSN6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COPS6 expression without altering the underlying DNA sequence.

CSN6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COPS6 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 COPS6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CSN6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COPS6 locus and enabling the study of CSN6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CSN6 pathway restoration in tumor cells with silenced or reduced COPS6 expression.

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