The DUOXA1 Knockout HAP1 Polyclonal Cells are a genetically engineered human cell population with disruption of the DUOXA1 gene by CRISPR/Cas9-mediated gene editing. This polyclonal knockout model provides a heterogeneous pool of cells carrying diverse loss-of-function mutations, enabling robust functional studies without clonal selection bias. The cells serve as a ready-to-use tool for investigating DUOXA1-dependent redox signaling, innate immunity, and thyroid hormone biosynthesis in a well-defined genomic background.
The host cell line, HAP1, is a near-haploid chronic myeloid leukemia (CML) line derived from KBM-7. Its haploid karyotype allows complete gene knockout with a single allelic inactivation, eliminating heterozygous compensation. HAP1 cells are widely used in functional genomics and genetic screens due to their stable growth, ease of transfection, and simplified genetic background. They provide a clean system for studying oxidative stress pathways, as their basal redox state is well characterized.
DUOXA1 is an essential chaperone that facilitates the maturation and membrane localization of the dual oxidases DUOX1 and DUOX2. These enzymes produce extracellular hydrogen peroxide (H2O2) critical for epithelial host defense and thyroid hormone synthesis. DUOXA1 expression is induced by inflammatory cytokines such as TNF-?? and IL-4/IL-13 via the transcription factors NF-??B and STAT6. It forms complexes with DUOX1/2 and ER chaperones like calnexin, ensuring proper protein folding. Downstream, H2O2 serves as substrate for lactoperoxidase-mediated antimicrobial activity and thyroid peroxidase-catalyzed iodination.
In the HAP1 context, DUOXA1 knockout provides a unique platform to study redox signaling in a leukemia-derived background. The haploid nature ensures complete loss of DUOXA1 function, enabling clean dissection of its role in DUOX enzyme regulation. Although HAP1 cells are not classical epithelial cells, they can be used to reconstitute the DUOX system for mechanistic studies. This model is also valuable for exploring the role of ROS modulation in cancer cell survival and for screening inhibitors of the DUOX pathway.
Research applications include quantifying H2O2 production via Amplex Red assays, assessing DUOX1/2 surface expression by flow cytometry, and measuring thyroid peroxidase activity. The cells are suitable for antimicrobial killing assays and high-throughput screens for DUOX modulators. In cancer biology, they enable investigation of redox-dependent synthetic lethalities. For further details, contact Ascent Research.
