ZIP2 CRISPR Activation Plasmid (h): sc-412062-ACT

SLC39A2 encodes ZIP2, a plasma membrane zinc influx transporter that regulates cytosolic Zn2+ availability and supports zinc-dependent enzymatic activity, protein folding, and transcriptional control. By shaping zinc homeostasis, ZIP2 influences processes such as epithelial differentiation, oxidative stress responses, and signal transduction pathways that rely on zinc-sensitive regulators. Altered zinc transport has been associated with disrupted barrier function and inflammatory signaling, linking ZIP2 biology to contexts including epithelial dysfunction and cancer-related phenotypes. As a component of the broader ZIP/SLC39 network, ZIP2 provides a tractable node for dissecting how zinc flux modulates cellular metabolism and stress-adaptive programs.

ZIP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC39A2 expression without altering the underlying DNA sequence.

ZIP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC39A2 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 SLC39A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZIP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC39A2 locus and enabling the study of ZIP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZIP2 pathway restoration in tumor cells with silenced or reduced SLC39A2 expression.

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