The EBI3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in the near-haploid HAP1 cell line, targeting the human EBI3 gene. This product provides a loss-of-function model to investigate the roles of EBI3 in immune signaling networks. The polyclonal format offers a heterogeneous population of edited cells, facilitating robust functional studies without clonal selection biases.
The HAP1 host cell line is derived from the KBM-7 chronic myeloid leukemia cell line and displays a near-haploid karyotype, except for disomy of chromosome 8. Originating from a male donor, HAP1 cells carry the Philadelphia chromosome and are widely employed in genetic screens, CRISPR-based functional genomics, and targeted gene studies due to their haploid nature, which simplifies gene disruption and genotype?Cphenotype analyses.
EBI3 encodes the beta subunit of the heterodimeric cytokines interleukin-27 (IL-27) and interleukin-35 (IL-35). IL-27, composed of EBI3 and p28 (IL-27A), binds the IL-27 receptor (WSX-1/gp130) to activate JAK1/JAK2 and STAT1/STAT3 signaling. This pathway is triggered by upstream regulators such as TLR4 ligands (LPS), IFN-gamma, and CD40 ligation, and leads to the induction of T-bet and IL-10, promoting Th1 differentiation and anti-inflammatory responses. IL-35, formed by EBI3 and IL-12p35, interacts with a receptor consisting of gp130 and IL-12R??2, activating STAT1/STAT4 and driving the expansion of Foxp3+ regulatory T cells (Tregs) while suppressing Th17 cell development. Thus, EBI3 sits at the crossroads of pro- and anti-inflammatory signaling.
In the HAP1 background, knockout of EBI3 disrupts both IL-27 and IL-35 cytokine assembly, providing a clean system to parse their distinct contributions to immune regulation. The near-haploid genotype reduces genetic redundancy, enabling clear interrogation of EBI3-dependent pathways. This model is particularly valuable for high-throughput screens designed to identify modifiers of IL-27/IL-35 signaling or to validate therapeutic targets in immune-related diseases.
Typical research applications include mechanistic studies of IL-27 and IL-35 signaling, T cell differentiation and immune tolerance, cancer immunology, and autoimmune disease modeling. The EBI3 knockout HAP1 polyclonal cells support assays such as western blotting for knockout confirmation, ELISA for cytokine secretion, flow cytometry for T cell subset analysis, phospho-STAT detection, RT-qPCR for target gene expression, T cell functional assays, RNA-seq, and co-immunoprecipitation of EBI3 with p28 or p35. For further information, please contact Ascent Research.
