ZDHHC5 CRISPR Activation Plasmid (h): sc-411833-ACT

ZDHHC5 encodes a DHHC-family palmitoyltransferase that catalyzes S-palmitoylation of membrane-associated proteins, regulating their trafficking, stability, and compartmentalization at the plasma membrane and endosomal systems. Through control of dynamic lipidation cycles, ZDHHC5 influences signal transduction, synaptic and immune receptor organization, and cytoskeletal remodeling, with downstream effects on cell adhesion and migration. Dysregulated palmitoylation networks involving ZDHHC5 have been linked to altered neuronal signaling and oncogenic pathway modulation, making it relevant for studies of neurobiology and cancer cell behavior. As a membrane-localized enzyme, ZDHHC5 is frequently investigated in the context of protein localization, vesicular transport, and post-translational modification-dependent signaling.

ZDHHC5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZDHHC5 expression without altering the underlying DNA sequence.

ZDHHC5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZDHHC5 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 ZDHHC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZDHHC5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZDHHC5 locus and enabling the study of ZDHHC5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZDHHC5 pathway restoration in tumor cells with silenced or reduced ZDHHC5 expression.

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