SENP5 CRISPR Activation Plasmid (h): sc-404428-ACT

Human SENP5 encodes a SUMO-specific protease that catalyzes deSUMOylation and SUMO precursor processing, helping maintain SUMO homeostasis across nuclear and mitochondrial-associated processes. By reversing SUMO conjugation on key substrates, SENP5 influences cell-cycle progression, mitotic spindle organization, DNA damage responses, and stress-adaptive transcriptional programs. SENP5 activity is linked to regulation of protein stability and subcellular localization through the SUMO pathway, intersecting with ubiquitin-dependent turnover and chromatin-associated signaling. Dysregulated SUMO protease function, including altered SENP5 expression or activity, has been associated with aberrant proliferation, genomic instability, and cancer-relevant signaling phenotypes in experimental models.

SENP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SENP5 expression without altering the underlying DNA sequence.

SENP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SENP5 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 SENP5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SENP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SENP5 locus and enabling the study of SENP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SENP5 pathway restoration in tumor cells with silenced or reduced SENP5 expression.

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