The IL3 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HAP1 human near-haploid cell line. This pool of edited cells provides a robust model for analyzing IL3-dependent signaling pathways and cellular responses. By targeting the IL3 locus with CRISPR/Cas9, the population offers a powerful tool for functional genomics and cytokine biology research.
HAP1 cells are a near-haploid derivative of the KBM-7 chronic myeloid leukemia (CML) line, harboring the BCR-ABL fusion oncogene. They grow in suspension as hematopoietic cells and are prized for functional genomics studies due to their simplified genome. This background allows efficient gene targeting and minimizes genetic redundancy, making HAP1 an optimal host for knockout modeling in leukemic contexts.
Interleukin-3 (IL3) is a hematopoietic growth factor secreted by activated T cells and other immune cells. It binds the heterodimeric receptor composed of IL3RA and CSF2RB, triggering phosphorylation of JAK2 and subsequent activation of STAT5, as well as the MAPK/ERK and PI3K/AKT pathways. Downstream, these signals induce expression of anti-apoptotic BCL2, the transcription factor c-MYC, and cyclin D1 (CCND1) to drive cell-cycle entry and survival. Upstream regulators of IL3 include TCR activation, NFAT transcription factors, IL2, and GM-CSF, placing IL3 at a critical node in hematopoietic cytokine networks.
In the BCR-ABL-positive HAP1 background, IL3 knockout enables dissection of oncogene-driven versus cytokine-driven growth mechanisms. Loss of IL3 impairs JAK2/STAT5 and MAPK/ERK signaling, reducing BCL2 and cyclin D1 levels and thereby sensitizing cells to apoptosis. This model is particularly informative for investigating how autocrine IL3 loops support leukemic cell survival and for testing combinations of cytokine receptor antagonists with existing kinase inhibitors.
Applications include cytokine signaling studies, leukemia cell biology, functional genomic screens, and drug target validation. Typical assays include western blotting for phospho-STAT5, cell proliferation measurements, RT-qPCR of BCL2 and c-MYC transcripts, Annexin V apoptosis flow cytometry, and phospho-signaling analysis. These readouts enable thorough interrogation of IL3-mediated processes. For further information, please contact Ascent Research.
