DZIP3 CRISPR Activation Plasmid (h): sc-403972-ACT

Human DZIP3 encodes a RING-type E3 ubiquitin ligase implicated in ubiquitin-dependent protein turnover and regulation of signal transduction at the cellular membrane and cytoskeleton interface. Through selective substrate ubiquitination, DZIP3 can influence proteostasis, trafficking, and cell-state transitions that depend on coordinated remodeling of protein complexes. Dysregulation of ubiquitin–proteasome pathway components, including E3 ligases, is frequently associated with altered proliferation, differentiation, and stress-response programs relevant to cancer and neurobiology research. DZIP3 is therefore of interest for dissecting ubiquitin-mediated control of transcriptional outputs and pathway responsiveness in human cell models.

DZIP3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DZIP3 expression without altering the underlying DNA sequence.

DZIP3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DZIP3 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 DZIP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DZIP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DZIP3 locus and enabling the study of DZIP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DZIP3 pathway restoration in tumor cells with silenced or reduced DZIP3 expression.

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