ZDHHC2 CRISPR Activation Plasmid (h): sc-403195-ACT

ZDHHC2 encodes a DHHC family palmitoyltransferase that catalyzes S-palmitoylation of target proteins, modulating their membrane association, stability, trafficking, and signaling competence. Through dynamic palmitoylation cycles, ZDHHC2 contributes to regulation of vesicular transport, synaptic and immune-related signaling, and organization of membrane microdomains that influence downstream pathway output. Altered ZDHHC2 activity and palmitoylation homeostasis have been associated with dysregulated cell signaling networks implicated in cancer biology and neurological processes, making it a useful entry point for studying how lipid modifications tune protein function. As a human enzyme acting at membranes, ZDHHC2 is frequently investigated alongside palmitoylation/depalmitoylation machinery and pathway-specific substrates to map functional consequences of post-translational lipidation.

ZDHHC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZDHHC2 expression without altering the underlying DNA sequence.

ZDHHC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZDHHC2 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 ZDHHC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZDHHC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZDHHC2 locus and enabling the study of ZDHHC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZDHHC2 pathway restoration in tumor cells with silenced or reduced ZDHHC2 expression.

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