gankyrin CRISPR Activation Plasmid (h): sc-402164-ACT

PSMD10 encodes gankyrin, an ankyrin repeat–containing oncoprotein that associates with the 26S proteasome regulatory complex and modulates ubiquitin-dependent protein turnover. By interacting with key tumor suppressor pathways, including RB1/E2F and p53 signaling, gankyrin influences cell-cycle progression, checkpoint control, and apoptosis. Aberrant PSMD10 expression has been linked to dysregulated proliferation and altered proteostasis in multiple cancer contexts, supporting its use as a mechanistic node for studying transcriptional control and protein degradation networks. In addition, gankyrin impacts stress-response and inflammatory signaling through effects on protein stability and regulatory complex assembly.

gankyrin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMD10 expression without altering the underlying DNA sequence.

gankyrin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMD10 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 PSMD10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous gankyrin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMD10 locus and enabling the study of gankyrin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of gankyrin pathway restoration in tumor cells with silenced or reduced PSMD10 expression.

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