ZnT-4 CRISPR Activation Plasmid (h): sc-403951-ACT

SLC30A4 encodes ZnT-4 (SLC30 family), a zinc efflux transporter that regulates cytosolic zinc by promoting zinc sequestration into intracellular compartments, thereby shaping zinc-dependent enzymatic activity and signaling. By tuning zinc availability, ZnT-4 contributes to metal ion homeostasis, vesicular trafficking, and redox balance, processes that intersect with pathways controlling oxidative stress responses and secretory function. Dysregulated zinc transport has been linked broadly to altered cellular differentiation, impaired secretory granule biology, and inflammatory and metabolic phenotypes, making SLC30A4 a useful locus for mechanistic studies of zinc-dependent cell physiology. Research on ZnT-4 supports investigations into how compartmentalized zinc influences transcriptional programs and stress-adaptive signaling in human cells.

ZnT-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC30A4 expression without altering the underlying DNA sequence.

ZnT-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC30A4 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 SLC30A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZnT-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC30A4 locus and enabling the study of ZnT-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZnT-4 pathway restoration in tumor cells with silenced or reduced SLC30A4 expression.

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