PSMC4 CRISPR Activation Plasmid (h): sc-403885-ACT

PSMC4 encodes an AAA+ ATPase subunit of the 19S regulatory particle of the 26S proteasome, supporting recognition, unfolding, and translocation of polyubiquitinated substrates into the catalytic core. Through this role in the ubiquitin–proteasome system, PSMC4 contributes to proteostasis, turnover of cell-cycle regulators, modulation of stress responses, and control of transcriptional programs influenced by protein stability. Proteasome activity intersects with pathways such as NF-κB signaling, DNA damage responses, and adaptive responses to proteotoxic stress, making PSMC4 relevant to studies of proliferation and cellular homeostasis. Dysregulated proteasome regulation and altered expression of proteasome subunits have been associated with oncogenic states and neurodegeneration-related proteostasis defects, supporting investigation of PSMC4 in disease-relevant cellular models.

PSMC4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMC4 expression without altering the underlying DNA sequence.

PSMC4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMC4 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 PSMC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSMC4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMC4 locus and enabling the study of PSMC4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSMC4 pathway restoration in tumor cells with silenced or reduced PSMC4 expression.

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