The HMOX2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of HeLa cells with targeted disruption of the human HMOX2 gene, resulting in loss of constitutive heme oxygenase-2 (HO-2) function. This polyclonal model captures a spectrum of gene-editing events, providing a robust system for studying HO-2-dependent cellular processes without relying on single-cell clonal isolation. Loss of HO-2 activity eliminates the primary constitutive source of carbon monoxide and biliverdin/bilirubin, allowing dissection of heme catabolism and redox signaling.
HeLa cells, an epithelial line derived from HPV18-positive cervical adenocarcinoma, are a cornerstone of biomedical research due to their rapid growth, stability, and extensive characterization. Engineered for HMOX2 knockout, this HeLa derivative enables investigation of constitutive heme oxygenase function within a well-studied cancer cell context, facilitating studies on cytoprotection, oxidative stress, and iron homeostasis.
HMOX2 encodes the constitutive heme oxygenase isoform that cleaves heme into biliverdin, carbon monoxide (CO), and free iron (Fe2?). Biliverdin is reduced to the antioxidant bilirubin by biliverdin reductase, while CO activates soluble guanylyl cyclase to produce cGMP, triggering protein kinase G-mediated protective pathways. HO-2 activity is modulated by heme, nitric oxide, casein kinase 2, and oxidative stress, and requires electron transfer from NADPH-cytochrome P450 reductase and cytochrome b5. Downstream, HO-2 influences iron sequestration, redox balance, and anti-apoptotic signaling, positioning it as a key player in cellular resilience.
In HeLa cells, HMOX2 knockout abrogates constitutive CO and bilirubin production, impairing antioxidant defenses and potentially sensitizing cells to oxidative damage. This model is valuable for exploring how cancer cells manage intrinsic oxidative stress and for testing the role of HO-2 in proliferation, chemoresistance, and ferroptosis. The polyclonal nature minimizes clonal bias and better mimics heterogeneous tumor populations, enhancing physiological relevance.
Applications include heme oxygenase activity assays, CO and bilirubin detection, intracellular iron measurement with calcein-AM, and ROS detection using H2DCFDA or MitoSOX. Western blotting and RT-qPCR can profile stress-response proteins, while viability assays under oxidative challenge assess cytoprotection. The cells are suitable for drug screening targeting heme oxygenase modulators and for dissecting CO/cGMP signaling. For further details, contact Ascent Research.
