SUMO-2 CRISPR Activation Plasmid (h): sc-401111-ACT

Human SUMO2 encodes SUMO-2, a ubiquitin-like modifier that is covalently conjugated to target proteins in a dynamic post-translational modification cycle regulating protein stability, subcellular localization, and macromolecular complex assembly. SUMO-2–dependent SUMOylation influences DNA damage signaling and repair, chromatin organization, transcriptional control, and cell-cycle progression through SUMO E1/E2/E3 enzyme cascades and coordinated deSUMOylation by SENP proteases. SUMO-2 can form poly-SUMO chains that interface with SUMO-targeted ubiquitin ligases, linking SUMOylation to proteostasis and stress-adaptive responses. Dysregulated SUMO2 activity and altered SUMOylation patterns are frequently studied in contexts of oncogenic signaling, neurodegeneration, and inflammatory stress where network-level changes in transcription and genome maintenance are implicated.

SUMO-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SUMO2 expression without altering the underlying DNA sequence.

SUMO-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SUMO2 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 SUMO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SUMO-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SUMO2 locus and enabling the study of SUMO-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SUMO-2 pathway restoration in tumor cells with silenced or reduced SUMO2 expression.

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