UBE2S CRISPR Activation Plasmid (h): sc-404140-ACT

UBE2S (ubiquitin-conjugating enzyme E2 S) is an E2 ubiquitin–conjugating enzyme that extends K11-linked polyubiquitin chains, working with the anaphase-promoting complex/cyclosome (APC/C) to promote timely ubiquitination and degradation of cell-cycle regulators. Through this activity, UBE2S helps coordinate mitotic progression, spindle checkpoint satisfaction, and orderly exit from mitosis, thereby supporting genome stability and controlled proliferation. Dysregulated UBE2S expression or activity has been associated with aberrant proteostasis and cell-cycle control in multiple disease contexts, including cancers where ubiquitin pathway remodeling is common. As a node in ubiquitin-dependent signaling, UBE2S is also used to study crosstalk between APC/C function, proteasome-mediated turnover, and stress-responsive pathways that influence cell fate decisions.

UBE2S CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UBE2S expression without altering the underlying DNA sequence.

UBE2S CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UBE2S 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 UBE2S transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UBE2S expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UBE2S locus and enabling the study of UBE2S-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UBE2S pathway restoration in tumor cells with silenced or reduced UBE2S expression.

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