DUOXA2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DUOXA2 gene in the near-haploid human HAP1 cell line. This genetically heterogeneous pool of edited cells enables loss-of-function studies of DUOXA2 without the clonal bias inherent to single-cell-derived lines. The product provides a physiologically relevant model for investigating DUOXA2-dependent processes in thyroid hormone biosynthesis and reactive oxygen species (ROS) signaling.
The HAP1 host cell line is derived from KBM-7 chronic myeloid leukemia cells and exhibits a near-haploid karyotype, making it an ideal platform for genetic screening and knockout studies. Its haploid nature simplifies functional genomics, as disruption of a single allele is sufficient to produce a null phenotype, eliminating the complexity of diploid compensation. This background is widely employed in both arrayed and pooled CRISPR screens to identify genetic dependencies, validate gene function, and dissect signaling pathways.
DUOXA2 encodes an endoplasmic reticulum chaperone that is specifically required for the maturation and cell surface expression of DUOX2, the dual oxidase responsible for generating hydrogen peroxide (H2O2) in the thyroid gland. DUOXA2 expression is transcriptionally regulated by thyroid transcription factors PAX8 and NKX2-1, which act downstream of TSH receptor (TSHR) signaling via the cAMP/PKA cascade. The DUOXA2 protein forms a stable complex with DUOX2, facilitating its proper folding and trafficking. Functional DUOX2 at the apical membrane produces H2O2, which serves as a co-substrate for thyroid peroxidase (TPO)-mediated iodination of thyroglobulin (TG); iodide is actively imported by the sodium-iodide symporter (NIS, SLC5A5). This pathway is essential for the synthesis of thyroid hormones T4 and T3, and its disruption is linked to congenital hypothyroidism and thyroid dyshormonogenesis type 5.
In the HAP1 background, polyclonal knockout of DUOXA2 offers a robust system to dissect the redox-dependent machinery of thyroid hormone production without the artifacts of clonal selection. The near-haploid state ensures straightforward genotype-phenotype correlations, while the polyclonal nature maintains population-level heterogeneity that can better model tissue-level responses. Researchers can use these cells to explore how DUOXA2 loss impacts DUOX2 stability, H2O2 generation, and downstream iodination events, providing insights into ER chaperone function and oxidative stress physiology.
Typical applications include modeling congenital hypothyroidism associated with DUOXA2 mutations, investigating the chaperone-dependent regulation of DUOX2, screening for genetic modifiers of ROS production, and performing detailed mechanistic studies of thyroid hormone biosynthesis. These knockout cells are compatible with a range of experimental techniques, such as Western blotting and RT-qPCR for expression analysis, immunofluorescence and flow cytometry to assess DUOX2 surface localization, Amplex Red assays for H2O2 quantification, co-immunoprecipitation for DUOXA2?CDUOX2 interaction, and iodide uptake measurements to evaluate organification. For further details or technical support, please contact Ascent Research.
