Mms2 CRISPR Activation Plasmid (h): sc-404777-ACT

UBE2V2 encodes Mms2, an E2 ubiquitin-conjugating enzyme variant that lacks catalytic activity but forms a functional heterodimer with UBE2N/UBC13 to promote K63-linked polyubiquitin chain assembly. This non-proteolytic ubiquitin signaling supports DNA damage tolerance and repair, including post-replication repair and error-free lesion bypass mediated by PCNA ubiquitination. Mms2-dependent ubiquitin chain formation also contributes to stress-responsive signaling networks that intersect with genome stability and cell-cycle checkpoint control. Dysregulation of ubiquitin-mediated DNA repair pathways involving UBE2V2 has been associated with altered mutational burden and cellular sensitivity to genotoxic stress, making it relevant for mechanistic studies in cancer biology and DNA repair disorders.

Mms2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UBE2V2 expression without altering the underlying DNA sequence.

Mms2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UBE2V2 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 UBE2V2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mms2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UBE2V2 locus and enabling the study of Mms2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mms2 pathway restoration in tumor cells with silenced or reduced UBE2V2 expression.

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