PSMD1 CRISPR Activation Plasmid (h): sc-405171-ACT

PSMD1 encodes a non-ATPase subunit of the 19S regulatory particle of the 26S proteasome, a central machinery for ATP-dependent degradation of polyubiquitinated proteins. By contributing to substrate recognition and proteasome assembly, PSMD1 supports protein quality control, cell-cycle progression, DNA damage responses, and stress-adaptive signaling through ubiquitin–proteasome system pathways. Proteasome regulatory subunits influence turnover of key pathway mediators, including components of NF-κB signaling and other transcriptional programs, linking PSMD1 activity to inflammatory and proliferative states. Dysregulated proteasome function and altered expression of proteasome subunits are frequently associated with oncogenic phenotypes, proteotoxic stress, and neurodegeneration-relevant protein homeostasis defects.

PSMD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMD1 expression without altering the underlying DNA sequence.

PSMD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMD1 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 PSMD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSMD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMD1 locus and enabling the study of PSMD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSMD1 pathway restoration in tumor cells with silenced or reduced PSMD1 expression.

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