This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human IRAK1 gene in the HAP1 near-haploid cell line. The IRAK1 Knockout HAP1 Polyclonal Cells provide a loss-of-function model for interleukin-1 receptor-associated kinase 1 (IRAK1), a key serine/threonine kinase in innate immune signaling. The polyclonal format yields a heterogeneous pool of edited alleles, enabling robust functional studies without clonal selection bias.
HAP1 cells are a near-haploid human male cell line derived from chronic myeloid leukemia KBM-7. Their haploid karyotype simplifies genetic manipulation and phenotype interpretation, making HAP1 an established model for genetic screens, drug target validation, and cancer research. The near-haploid background reduces confounding effects from second alleles, facilitating clean functional analyses.
IRAK1 functions downstream of Toll-like receptors (TLRs) and interleukin-1 receptor (IL-1R) as an adaptor kinase in the MYD88-dependent pathway. Upon stimulation, IRAK1 is recruited to the receptor complex via MYD88 and IRAK4, leading to its phosphorylation. Active IRAK1 associates with TRAF6, promoting TAK1-mediated activation of the IKK complex and MAP kinases, which drive NF-??B and AP-1 transcription factors. This induces expression of pro-inflammatory cytokines (TNF, IL-6, IL-1??). Regulatory interactions with Pellino-1 and TAB2 modulate signaling.
Disruption of IRAK1 in HAP1 creates a powerful system for dissecting innate immune pathways and their disease implications. The knockout allows examination of TLR/IL-1R signaling defects and compensatory mechanisms. Given HAP1??s CML origin, this model also supports investigation of inflammatory signaling in hematologic malignancies. Applications span from basic discovery in rheumatoid arthritis and lupus to target validation in cancer and inflammation. These cells enable detailed analysis of IRAK1-dependent cytokine production and serve as a platform for testing small-molecule inhibitors of upstream or downstream pathway components.
Typical assays include western blotting and RT-qPCR for expression analysis, NF-??B luciferase reporter assays, co-immunoprecipitation for protein interactions, and cytokine profiling by ELISA or bead arrays. Phospho-kinase arrays and flow cytometry assess pathway activation, while RNA-seq and proteomics enable global profiling. Researchers may also perform functional rescue experiments by re-expressing wild-type or mutant IRAK1 alleles to validate specific signaling functions. These cells are suitable for kinase inhibitor screens and pathway interrogation. For further information, please contact Ascent Research.
