ZIP4 CRISPR Activation Plasmid (h): sc-412558-ACT

SLC39A4 encodes the human zinc transporter ZIP4, a multi-pass membrane protein that mediates cellular uptake of extracellular Zn2+ and helps maintain zinc-dependent metabolic and signaling homeostasis. ZIP4 activity supports processes including epithelial differentiation, barrier integrity, and metal-responsive transcriptional programs, with downstream effects on oxidative stress responses and protein synthesis pathways that require zinc as a structural or catalytic cofactor. Dysregulated SLC39A4 expression or function perturbs zinc availability and has been linked to disorders of zinc absorption and broader phenotypes involving growth, immune function, and epithelial tissue maintenance. Because zinc influences numerous enzymes and transcription factors, ZIP4 is frequently studied as an upstream regulator of nutrient-sensing and stress-adaptation networks.

ZIP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC39A4 expression without altering the underlying DNA sequence.

ZIP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC39A4 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 SLC39A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZIP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC39A4 locus and enabling the study of ZIP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZIP4 pathway restoration in tumor cells with silenced or reduced SLC39A4 expression.

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