SUMO-3 CRISPR Activation Plasmid (m): sc-423045-ACT

Mouse Sumo3 encodes SUMO-3, a ubiquitin-like modifier that is covalently conjugated to lysine residues on target proteins to regulate their stability, localization, and activity. SUMOylation coordinates dynamic control of transcription, DNA damage repair, chromatin organization, and cell-cycle progression through the E1 (SAE1/SAE2), E2 (UBC9), and E3 ligase network, and is reversed by SENP proteases. SUMO-3 participates in stress-responsive signaling and proteostasis by modulating nuclear body assembly and protein–protein interaction networks. Dysregulated SUMO conjugation has been linked in the literature to altered inflammatory signaling, neurodegeneration-associated proteotoxic stress, and oncogenic transcriptional programs, supporting its use in mechanistic disease biology studies.

SUMO-3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sumo3 expression without altering the underlying DNA sequence.

SUMO-3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sumo3 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 Sumo3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SUMO-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sumo3 locus and enabling the study of SUMO-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SUMO-3 pathway restoration in tumor cells with silenced or reduced Sumo3 expression.

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