ZDHHC20 CRISPR Activation Plasmid (h): sc-406353-ACT

ZDHHC20 encodes a DHHC family palmitoyltransferase that catalyzes S-palmitoylation of cytosolic cysteine residues on membrane-associated proteins, shaping their trafficking, stability, and signaling competence. By modulating dynamic palmitoylation cycles, ZDHHC20 influences organization of membrane microdomains and downstream pathways linked to receptor signaling, vesicular transport, and cellular stress responses. Altered palmitoylation homeostasis has been associated with dysregulated growth and immune signaling, and ZDHHC20 has been investigated in the context of cancer biology, metabolic regulation, and host–pathogen interactions. These features make ZDHHC20 a useful node for dissecting post-translational lipidation mechanisms and pathway rewiring in human cells.

ZDHHC20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZDHHC20 expression without altering the underlying DNA sequence.

ZDHHC20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZDHHC20 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 ZDHHC20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZDHHC20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZDHHC20 locus and enabling the study of ZDHHC20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZDHHC20 pathway restoration in tumor cells with silenced or reduced ZDHHC20 expression.

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