p40-phox CRISPR Activation Plasmid (h): sc-401461-ACT

NCF4 encodes p40-phox, a regulatory component of the phagocyte NADPH oxidase (NOX2) complex that modulates assembly and activity of the oxidase on endomembranes and phagosomes. Through interactions with p67-phox (NCF2), p47-phox (NCF1), and membrane cytochrome b558, p40-phox helps control stimulus-dependent reactive oxygen species generation that supports microbial killing and redox signaling. This pathway integrates with innate immune processes including phagocytosis, inflammasome signaling, and cytokine responses, where altered oxidase regulation can perturb host defense and inflammatory homeostasis. Dysregulated NOX2 activity and p40-phox-dependent signaling have been linked to immunodeficiency and inflammatory phenotypes, providing a mechanistic entry point for studying oxidative burst regulation in myeloid cells.

p40-phox CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCF4 expression without altering the underlying DNA sequence.

p40-phox CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCF4 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 NCF4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p40-phox expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCF4 locus and enabling the study of p40-phox-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p40-phox pathway restoration in tumor cells with silenced or reduced NCF4 expression.

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