JAB1 CRISPR Activation Plasmid (m): sc-424107-ACT

Mouse Cops5 encodes JAB1/CSN5, the catalytically active deneddylase subunit of the COP9 signalosome that regulates cullin-RING ubiquitin ligases and proteostasis. By controlling NEDD8 removal from cullins, JAB1 modulates ubiquitin-dependent turnover of key cell-cycle and signaling regulators, influencing proliferation, DNA damage responses, and stress-adaptive programs. JAB1 also interfaces with transcriptional control and signal transduction networks, including pathways linked to AP-1 activity and cytokine-responsive signaling. Dysregulated CSN5/JAB1 activity has been associated with aberrant cell growth and genomic instability in cancer biology, making it a useful node for mechanistic studies of oncogenic signaling and protein homeostasis.

JAB1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cops5 expression without altering the underlying DNA sequence.

JAB1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cops5 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 Cops5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JAB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cops5 locus and enabling the study of JAB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JAB1 pathway restoration in tumor cells with silenced or reduced Cops5 expression.

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