MPO/Myeloperoxidase CRISPR Activation Plasmid (h): sc-400822-ACT

Human MPO encodes myeloperoxidase, a heme-containing peroxidase highly expressed in neutrophils and monocytes that catalyzes the conversion of hydrogen peroxide and halides into hypohalous acids, including hypochlorous acid. This oxidative chemistry supports innate immune defense, neutrophil extracellular trap formation, and modulation of redox-sensitive signaling that influences inflammation and tissue remodeling. MPO activity intersects with reactive oxygen species pathways, phagosome maturation, and host–pathogen interactions, and can also promote oxidative modification of proteins and lipids. Dysregulated MPO expression or activity has been associated with inflammatory disorders, cardiovascular disease risk, and tumor-associated myeloid phenotypes, making it a useful target for mechanistic studies of immunometabolism and oxidative stress.

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

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

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