PSMC2 CRISPR Activation Plasmid (h): sc-405022-ACT

PSMC2 encodes the 26S proteasome regulatory subunit 7 (Rpt1), an AAA+ ATPase within the 19S regulatory particle that powers substrate unfolding and translocation into the 20S core for ubiquitin-dependent protein degradation. Through control of proteostasis, PSMC2 influences cell-cycle progression, DNA damage responses, stress signaling, and antigen processing, integrating with pathways such as ubiquitin–proteasome turnover, NF-κB regulation, and proteotoxic stress adaptation. Altered proteasome activity and regulatory subunit expression are frequently associated with proliferative phenotypes and dysregulated checkpoint control observed across diverse disease contexts, making PSMC2 a useful node for studying protein quality control and regulatory network remodeling. Perturbing PSMC2 levels can therefore help interrogate how ATP-dependent proteasome functions shape transcriptional programs and cellular fitness.

PSMC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSMC2 expression without altering the underlying DNA sequence.

PSMC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSMC2 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 PSMC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSMC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSMC2 locus and enabling the study of PSMC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSMC2 pathway restoration in tumor cells with silenced or reduced PSMC2 expression.

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