POH1 CRISPR Activation Plasmid (h): sc-405591-ACT

PSMD14 encodes POH1, a JAMM/MPN+ metalloprotease subunit of the 19S regulatory particle of the human 26S proteasome that removes K48-linked ubiquitin chains during substrate processing. By coupling deubiquitination to proteasomal degradation, POH1 helps maintain protein homeostasis and shapes turnover of key regulators involved in DNA damage responses, cell-cycle progression, and stress-adaptive signaling. PSMD14 activity interfaces with ubiquitin-dependent control of chromatin-associated processes and can influence pathways such as NF-κB signaling through regulated degradation of pathway components. Dysregulation of proteasome-associated deubiquitination has been linked to aberrant proteostasis and genome stability phenotypes relevant to cancer biology and neurodegeneration research.

POH1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMD14 expression without altering the underlying DNA sequence.

POH1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMD14 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 PSMD14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous POH1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMD14 locus and enabling the study of POH1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of POH1 pathway restoration in tumor cells with silenced or reduced PSMD14 expression.

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