Heme Oxygenase 1/HMOX1 CRISPR Activation Plasmid (h): sc-400157-ACT

HMOX1 (heme oxygenase 1) is an inducible stress-response enzyme that catalyzes heme degradation to biliverdin, free iron, and carbon monoxide, linking heme metabolism to cellular redox balance. Its expression is strongly regulated by oxidative and electrophilic stress through pathways including NRF2–KEAP1 and inflammatory signaling, and it modulates mitochondrial function, iron handling, and cytoprotective antioxidant programs. HMOX1 activity influences macrophage polarization, endothelial responses, and tissue adaptation to hypoxia and inflammation, making it a widely used marker and mechanistic node in stress biology. Dysregulated HMOX1 has been associated with cardiometabolic and neuroinflammatory processes, tumor microenvironment remodeling, and altered susceptibility to oxidative injury, supporting broad relevance in disease modeling and pathway interrogation.

Heme Oxygenase 1/HMOX1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HMOX1 expression without altering the underlying DNA sequence.

Heme Oxygenase 1/HMOX1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HMOX1 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 HMOX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Heme Oxygenase 1/HMOX1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HMOX1 locus and enabling the study of Heme Oxygenase 1/HMOX1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Heme Oxygenase 1/HMOX1 pathway restoration in tumor cells with silenced or reduced HMOX1 expression.

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