NRAMP 1 CRISPR Activation Plasmid (m): sc-421957-ACT

Slc11a1 encodes NRAMP1, a phagosomal membrane transporter that regulates divalent metal ion flux, including iron and manganese, within macrophages and other myeloid cells. By shaping metal availability and phagolysosomal conditions, NRAMP1 influences antimicrobial effector functions, oxidative stress handling, and downstream innate immune signaling programs that coordinate inflammatory responses. In mouse models, Slc11a1 variation has been linked to altered susceptibility to intracellular pathogens and to immune-driven pathology through effects on macrophage activation state and cytokine networks. These properties make NRAMP1 a useful entry point for studying host–pathogen interactions, metal homeostasis in phagocytes, and immunometabolic regulation.

NRAMP 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc11a1 expression without altering the underlying DNA sequence.

NRAMP 1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc11a1 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 Slc11a1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NRAMP 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc11a1 locus and enabling the study of NRAMP 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NRAMP 1 pathway restoration in tumor cells with silenced or reduced Slc11a1 expression.

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