GODZ CRISPR Activation Plasmid (h): sc-403970-ACT

ZDHHC3, also known as GODZ, encodes a DHHC-family palmitoyl acyltransferase that catalyzes S-palmitoylation of cytosolic cysteine residues on target proteins, regulating their membrane association, trafficking, stability, and signaling output. This lipid modification influences vesicular transport and compartmentalization of receptors and channels, thereby shaping synaptic and membrane-proximal signaling networks. Through its role in post-translational modification and proteostasis, ZDHHC3 has been linked to dysregulated cell communication and stress responses that are frequently examined in cancer and neurological disease biology. As a result, ZDHHC3 is commonly studied for how palmitoylation-dependent remodeling of membrane microdomains impacts pathway activation and cellular phenotypes.

GODZ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZDHHC3 expression without altering the underlying DNA sequence.

GODZ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZDHHC3 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 ZDHHC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GODZ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZDHHC3 locus and enabling the study of GODZ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GODZ pathway restoration in tumor cells with silenced or reduced ZDHHC3 expression.

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