20S Proteasome α7 CRISPR Activation Plasmid (h): sc-401897-ACT

PSMA7 encodes the 20S proteasome α7 subunit, an essential structural component of the 20S core particle that supports assembly and gating of the proteolytic chamber in the ubiquitin–proteasome system. By enabling ATP-dependent turnover of polyubiquitinated substrates via the 26S proteasome, PSMA7 contributes to proteostasis, cell-cycle progression, antigen processing for MHC class I presentation, and regulation of stress-responsive signaling. Proteasome activity intersects with NF-κB signaling through IκB degradation and influences protein quality control pathways linked to oxidative stress and unfolded protein responses. Dysregulation of proteasome subunits, including PSMA7, is frequently studied in contexts of altered proliferation, inflammatory signaling, and neurodegenerative protein aggregation models.

20S Proteasome α7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMA7 expression without altering the underlying DNA sequence.

20S Proteasome α7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMA7 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 PSMA7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 20S Proteasome α7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMA7 locus and enabling the study of 20S Proteasome α7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 20S Proteasome α7 pathway restoration in tumor cells with silenced or reduced PSMA7 expression.

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