UBE2G2 CRISPR Activation Plasmid (h): sc-404851-ACT

UBE2G2 (ubiquitin-conjugating enzyme E2 G2) is a human E2 ubiquitin–protein ligase that transfers ubiquitin from E1 enzymes to E3 ligases, enabling substrate ubiquitination and downstream proteasomal degradation. It plays a prominent role in ER-associated degradation (ERAD) and protein quality control, coordinating with membrane-anchored E3 complexes to clear misfolded or regulatory proteins and maintain proteostasis. Through these functions, UBE2G2 intersects with cellular stress responses, cell-cycle control, and signaling pathways that depend on ubiquitin-mediated turnover. Dysregulated ubiquitin–proteasome activity and altered ERAD capacity are frequently linked to cancer biology and neurodegeneration-relevant proteostasis defects, making UBE2G2 a useful node for mechanistic studies of protein homeostasis.

UBE2G2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UBE2G2 expression without altering the underlying DNA sequence.

UBE2G2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UBE2G2 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 UBE2G2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UBE2G2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UBE2G2 locus and enabling the study of UBE2G2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UBE2G2 pathway restoration in tumor cells with silenced or reduced UBE2G2 expression.

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