NRAMP 2 CRISPR Activation Plasmid (h): sc-418176-ACT

Human SLC11A2 encodes NRAMP 2 (also known as DMT1), a proton-coupled divalent metal transporter that mediates cellular uptake of ferrous iron and other transition metals across endosomal and plasma membranes. By controlling iron influx from transferrin-containing endosomes and dietary absorption pathways, NRAMP 2 helps maintain cytosolic iron availability for heme synthesis, mitochondrial respiration, and iron–sulfur cluster biogenesis while influencing redox balance. Perturbation of SLC11A2 activity alters iron homeostasis and oxidative stress signaling, with documented relevance to disorders of iron metabolism such as microcytic anemia and systemic iron overload phenotypes. NRAMP 2 function is also studied in the context of metal handling in macrophages and neuronal vulnerability linked to iron-dependent cellular stress.

NRAMP 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC11A2 expression without altering the underlying DNA sequence.

NRAMP 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC11A2 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 SLC11A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NRAMP 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC11A2 locus and enabling the study of NRAMP 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NRAMP 2 pathway restoration in tumor cells with silenced or reduced SLC11A2 expression.

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