ZUFSP CRISPR Activation Plasmid (h): sc-415124-ACT

Human ZUFSP encodes a zinc finger–containing deubiquitinating enzyme that regulates ubiquitin-dependent protein quality control by cleaving specific ubiquitin chain linkages. It contributes to proteostasis and stress-responsive signaling by remodeling ubiquitin marks that govern protein turnover, DNA damage responses, and organelle homeostasis. ZUFSP activity intersects with ubiquitin–proteasome system dynamics and can influence pathway outputs controlled by ubiquitination, including cell-cycle progression and genome stability programs. Dysregulated ubiquitin editing is broadly implicated in cancer biology and neurodegeneration, making ZUFSP a useful node for mechanistic studies of ubiquitin signaling in human cells.

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

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

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