EDA Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HAP1 cells in which the human EDA gene has been disrupted, generating a loss-of-function model. This product provides a heterogeneous pool of knockout cells, circumventing clonal selection artifacts and offering a robust platform for functional genomics and signaling studies in a near-haploid genetic background.
The HAP1 cell line is derived from the KBM-7 chronic myelogenous leukemia line of hematopoietic origin. It is characterized by an adherent growth mode and a stable near-haploid karyotype, which reduce genetic redundancy and facilitate the interpretation of knockout phenotypes in functional assays.
Ectodysplasin A (EDA) is a type II transmembrane protein of the TNF superfamily that plays a critical role in ectodermal appendage development. EDA binds to the receptor EDAR, triggering the recruitment of the adaptor EDARADD and the E3 ubiquitin ligase TRAF6. This leads to activation of the IKK complex and the canonical NF-??B pathway, resulting in the transcriptional induction of downstream morphogens such as SHH, CCND1, WNT ligands, FGF20, and SOSTDC1. These factors coordinate the morphogenesis of hair follicles, teeth, and sweat glands. Upstream, EDA expression is regulated by the transcription factors TP63 and DLX, integrating developmental signals. Crosstalk with WNT signaling further modulates epithelial-mesenchymal interactions.
Despite their hematopoietic derivation, HAP1 cells provide a simplified and genetically accessible background in which to investigate the EDA/EDAR/NF-??B signaling module. The near-haploid state ensures that knockout phenotypes are fully penetrant, making this polyclonal product ideal for high-throughput drug screening, reporter-based assays, and mechanistic studies of pathway dynamics.
This knockout model is tailored for research on X-linked hypohidrotic ectodermal dysplasia (XLHED), non-syndromic tooth agenesis, and other EDA-related disorders. Users can monitor NF-??B activation via luciferase reporter assays, quantify downstream target expression (e.g., SHH, CCND1) by qRT-PCR, or assess NF-??B nuclear translocation by immunofluorescence. Western blotting for phospho-p65 and functional assays for apoptosis and proliferation further support drug discovery and toxicology screening. For additional information, please contact Ascent Research.
