PSMD7 CRISPR Activation Plasmid (h): sc-405949-ACT

PSMD7 (proteasome 26S subunit, non-ATPase 7) is a core component of the 19S regulatory particle of the human 26S proteasome, contributing to ubiquitin-dependent protein turnover and maintenance of cellular proteostasis. By supporting recognition and processing of polyubiquitinated substrates, PSMD7 influences pathways linked to cell-cycle progression, stress responses, and protein quality control, including ER-associated degradation and proteasome-mediated signaling events. Altered proteasome regulatory subunit activity can reshape degradation of key regulatory proteins, impacting transcriptional programs and adaptive responses to proteotoxic stress. Dysregulation of ubiquitin–proteasome system components, including PSMD7, is frequently studied in the context of proliferative signaling, genome maintenance, and disease-associated proteostasis imbalance.

PSMD7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMD7 expression without altering the underlying DNA sequence.

PSMD7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMD7 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 PSMD7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSMD7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMD7 locus and enabling the study of PSMD7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSMD7 pathway restoration in tumor cells with silenced or reduced PSMD7 expression.

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